3.6.4.13: RNA helicase
This is an abbreviated version!
For detailed information about RNA helicase, go to the full flat file.
Word Map on EC 3.6.4.13
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3.6.4.13
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single-stranded
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strand
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viruses
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duplex
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fork
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retinoic
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nucleic
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chromatin
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acid-inducible
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mda5
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dsrnas
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ssdna
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rna-binding
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stall
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werner
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nonstructural
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bloom
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polyproteins
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dna-dependent
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primase
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exonuclease
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replisome
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spliceosome
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hexameric
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minichromosome
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differentiation-associated
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dengue
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g-quadruplexes
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eif4g
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single-molecule
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unwound
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replicase
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restart
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ifn-stimulated
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nonsense-mediated
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rpa
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rad51
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holliday
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polyi:c
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flavivirus
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tfiih
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exoribonuclease
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ifn-beta
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d-loops
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pigmentosum
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cap-dependent
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xeroderma
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positive-stranded
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pre-rrnas
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overhang
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pharmacology
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medicine
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drug development
- 3.6.4.13
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single-stranded
- strand
- viruses
- duplex
- fork
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retinoic
- nucleic
- chromatin
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acid-inducible
- mda5
- dsrnas
- ssdna
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rna-binding
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stall
-
werner
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nonstructural
-
bloom
- polyproteins
-
dna-dependent
- primase
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exonuclease
- replisome
-
spliceosome
-
hexameric
-
minichromosome
-
differentiation-associated
- dengue
-
g-quadruplexes
- eif4g
-
single-molecule
-
unwound
-
replicase
-
restart
-
ifn-stimulated
-
nonsense-mediated
- rpa
- rad51
-
holliday
-
polyi:c
- flavivirus
- tfiih
- exoribonuclease
-
ifn-beta
-
d-loops
- pigmentosum
-
cap-dependent
- xeroderma
-
positive-stranded
- pre-rrnas
-
overhang
- pharmacology
- medicine
- drug development
Reaction
Synonyms
1a NTPase/helicase, Aquarius, AtHELPS, ATP-dependent helicase, ATP-dependent RNA helicase, ATP-dependent RNA helicase DDX3X, ATP-dependent RNA helicase DDX5, ATP/dATP-dependent RNA helicase, ATPase, ATPase/helicase, ATPase/RNA helicase, AtRH3, AtRH7, bel, BmL3-helicase, BMV 1a protein, BRR2, Brr2 RNA helicase, Brr2p, Cbu_0670, ChlR1 helicase, cold-shock DEAD-box protein A, CrhR, CsdA, CshA, CTHT_0005780, CTHT_0009470, DBP2, DbpA, DDX17, DDX19, DDX19B, DDX21, DDX21 RNA helicase, DDX25, DDX27, DDX3, DDX3X, DDX3Y, DDX4, DDX5, DDX58, DeaD, DEAD box helicase, DEAD box RNA helicase, DEAD-box ATP-dependent RNA helicase CshA, DEAD-box helicase, DEAD-box protein DED1, DEAD-box RNA helicase, DEAD-box rRNA helicase, DEAH box protein 34, DEAH-box protein 2, DEAH-box RNA helicase, DEAH/RHA RNA helicase, DED1, DENV NS3H, DEx(H/D)RNA helicase, DExD/H box RNA helicase, DEXD/H-box RNA helicase, dexh helicase, DExH protein RNA helicase A, DExH/D-Box protein RNA helicase A, DExH/D-box RNA helicase, DHX34, DHX36, DHX8, DHX9, Dhx9/RNA helicase A, DNA/RNA-dependent ATPase, EhDEAD1, EhDEAD1 RNA helicase, eIF4A, eIF4A helicase, eIF4AIII, eukaryotic initiation factor eIF 4A, frequency-interacting RNA helicase, FRH, FRQ-interacting RNA helicase, gonadotropin-regulated testicular RNA helicase, GRTH, GRTH/DDX25, HCV NS3 helicase, HEL-1, helicase, helicase 1, helicase B, helicase/nucleoside triphosphatase, HRpA, IBP160, increased size exclusion limit2, intron-binding protein 160, ISE2, KOKV helicase, Lmo1722, mitochondrial DEAD-box RNA helicase, Mss116p, mtr4, Mtr4p, NA-helicase, non structural protein 3, non-structural 3, non-structural protein 3, non-structural protein 3 protein, nonstructural protein 3, NPH-II, NS3, NS3 ATPase/helicase, NS3 helicase, NS3 NTPase/helicase, NS3 protein, NTPase/helicase, nucleoside 5'-triphosphatase, nucleoside triphosphatase/helicase, nucleoside triphosphatase/RNA helicase and 5'-RNA triphosphatase, NWMN_1985, p54 RNA helicase, p68, p68 RNA helicase, P72, PH1280, pre-mRNA-splicing ATP-dependent RNA helicase 28, protein NS3, Prp28, Prp43, Prp5, RENT1, RH22, RHA, RhlB, RIG-I, Rm62, RNA DEAD-box helicase, RNA helicase, RNA helicase A, RNA helicase Aquarius, RNA helicase BELLE, RNA helicase CrhR, RNA helicase CsdA, RNA helicase CshA, RNA helicase DDX17, RNA helicase DDX27, RNA helicase DDX3, RNA helicase Ddx39, RNA helicase DDX3X, RNA helicase DDX5, RNA helicase DDX6, RNA helicase DeaD, RNA helicase DHX34, RNA helicase DHX8, RNA helicase DHX9, RNA helicase HEL-1, RNA helicase Hera, RNA helicase ISE2, RNA Helicase p68, RNA helicase RHAU, RNA helicase UPF1, RNA splicing effector, RNA-dependent ATPase, RNA-dependent NTPase/helicase, RNA-helicase, RTPase, SA1885, Ski2-like helicase, slr0083, SNRNP200, spliceosomal DEAH-box RNA helicase, spliceosomal RNA helicase, SpolvlgA, Supv3L1, TaRH1, TGBp1 NTPase/helicase domain, Tk-DeaD, Triticum aestivum RNA helicase, Upf1, UPF1_1, UPF1_2, Vasa, VRH1, YxiN, ZmRH3, ZmRH3A, ZmRH3B
ECTree
3.6.4.13 RNA helicaseAdvanced search results
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results (Engineering
Engineering on EC 3.6.4.13 - RNA helicase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A115C/D262C
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site-directed mutagenesis, the mutant shows activity, structure and substrate specificity similar to the wild-type
A115C/E224C
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site-directed mutagenesis, the mutant shows activity, structure and substrate specificity similar to the wild-type
A115C/S229C
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site-directed mutagenesis, the mutant shows activity, structure and substrate specificity similar to the wild-type
S108C/E224C
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site-directed mutagenesis, the mutant shows activity, structure and substrate specificity similar to the wild-type
S108C/S229C
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site-directed mutagenesis, the mutant shows activity, structure and substrate specificity similar to the wild-type
D755A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 90% reduced ATPase activity compared to the wild-type enzyme
F788A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 30% reduced ATPase activity compared to the wild-type enzyme
G781S
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 75% reduced ATPase activity compared to the wild-type enzyme
H903A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 45% reduced ATPase activity compared to the wild-type enzyme
K691A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 80% reduced ATPase activity compared to the wild-type enzyme
Q785A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 65% reduced ATPase activity compared to the wild-type enzyme
Q785E
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 75% reduced ATPase activity compared to the wild-type enzyme
R791A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 10% increased ATPase activity compared to the wild-type enzyme
R815L
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 60% increased ATPase activity compared to the wild-type enzyme
R938A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 45% reduced ATPase activity compared to the wild-type enzyme
S790A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 60% reduced ATPase activity compared to the wild-type enzyme
S790W
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows 70% reduced ATPase activity compared to the wild-type enzyme
T812A/Y813A
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site-directed mutagenesis, mutation in the conserved BMV 1a protein helicase motif, the mutant shows abolished RNA recruitment and RNA stabilization, and thus RNA replication function, but normal accumulation, localization, and 2apol recruitment, the mutant shows unaltered ATPase activity compared to the wild-type enzyme
K232A
K199A/T200A
H51A
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site-directed mutagenesis, the mutant has an inactivated protease domain showing enhanced RNA helicase compared to wild-type full-length enzyme
R184Q/K185N/R186G/K187N
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construction of the N-terminally truncated mutant NS3DELTA180 containing a mutated RNA substrate biding motif, the mutant shows reduced RTPase activity
D310H
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site-directed mutagenesis of the V motif, leads to altered enzyme activity, overview
D313H
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site-directed mutagenesis of the V motif, leads to altered enzyme activity, overview
H320D
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site-directed mutagenesis of the V motif, leads to altered enzyme activity, overview
Y383A
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site-directed mutagenesis, the mutation causes the formation of a higher order molecular weight species in binding of RNaseE by RhlB
H293A
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mutation results in a protein with a significantly higher level of ATPase in the absence of RNA. The mutant protein still unwinds RNA. In the presence of RNA, the H293A mutant hydrolyzes ATP slower than wild-type
M1708A
S1369R
S1369R/M1708A
S1369R/Y1702A
Y1702A
DELTA53-105
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a region within the N-terminus of RHAU, referred to as the RSM, interacts with human telomerase RNA
F192E
site-directed mutagenesis, the mutant shows slightly reduced activity compared with the wild-type enzyme. Upf1F192E definitely prefers to unwind a dsDNA than to translocate it, strand switching
K235A
site-directed mutagenesis of a conserved Lys residue in RNA binding domain dsRBD2, the mutation does not prevent purified full-length RNA helicase A from binding and unwinding duplex RNA in vitro, but efficiently inhibits RNA helicase A-stimulated HIV-1 RNA metabolism including the accumulation of viral mRNA and tRNALys3 annealing to viral RNA
K236A
site-directed mutagenesis of a conserved Lys residue in RNA binding domain dsRBD2, the mutation does not prevent purified full-length RNA helicase A from binding and unwinding duplex RNA in vitro, but efficiently inhibits RNA helicase A-stimulated HIV-1 RNA metabolism including the accumulation of viral mRNA and tRNALys3 annealing to viral RNA
K50R
site-directed mutagenesis, ATPase dead mutant, fails completely to unwind triplex substrates
K54A
site-directed mutagenesis of a conserved Lys residue in RNA binding domain dsRBD1, the mutation does not prevent purified full-length RNA helicase A from binding and unwinding duplex RNA in vitro, but efficiently inhibits RNA helicase A-stimulated HIV-1 RNA metabolism including the accumulation of viral mRNA and tRNALys3 annealing to viral RNA
K54A/K55A
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mutant exhibits relatively minor reduction in interaction with SNV PCE
K54A/K55A/K236E
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triple mutant shows a severe reduction in interaction with junD post-transcriptional control element (PCE) or SNV PCE compared with wild-type
K55A
site-directed mutagenesis of a conserved Lys residue in RNA binding domain dsRBD1, the mutation does not prevent purified full-length RNA helicase A from binding and unwinding duplex RNA in vitro, but efficiently inhibits RNA helicase A-stimulated HIV-1 RNA metabolism including the accumulation of viral mRNA and tRNALys3 annealing to viral RNA
K829A
site-directed mutagenesis, mutation of the invariant lysine residue from motif I involved in ATP binding and hydrolysis drastically reduced Aquarius's ability to bind ATP and ADP and to hydroxadlyze ATP, the mutant shows significantly reduced ATPase activity compared to wild-type enzyme
K897del
site-directed mutagenesis, the mutant fails in unwinding the DNA substrates
Y1196A
site-directed mutagenesis, mutation of a conserved aromatic residue located near the putative RNA-binding surface of the RecA2 inhibits Aquarius's RNA-unwinding activity without changing its RNA-binding and ATPase properties, a helicase-deficient mutant
Y593F
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expression of the mutant enzyme in SW620 cells leads to Snail repression, E-cadherin upregulation and vimentin repression
G199A
mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost
G460A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities
G463A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities
K200A
mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200D
PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200E
PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200H
PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200N
PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200Q
PCR-based mutagenesis, ATPase and RNA helicase activity lost
K200R
PCR-based mutagenesis, ATPase and RNA helicase activity lost
Q457A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, 80% reduction of ATPase activity, no RNA helicase activity
R458A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, 90% reduction of ATPase activity, no RNA helicase activity
R459A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities
R461A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no ATPase activity, no RNA helicase activity
R464A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no ATPase activity, no RNA helicase activity
T201A
mutation in WALKER A motif, PCR-based mutagenesis, ATPase and RNA helicase activity lost
V462A
mutation of residues of the arginine finger within the active sites of ATP hydrolysis, no effect on either ATPase or RNA-unwinding activities
R806H
site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
V142G
site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
R806H
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site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
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V142G
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site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
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R806H
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site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
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V142G
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site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
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R806H
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site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
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V142G
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site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
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R806H
-
site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
-
V142G
-
site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
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R806H
-
site-directed mutagenesis, the mutation resides in a positively charged surface of the KOW domain, far removed from the helicase core, and disrupts circadian rhythms
-
V142G
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site-directed mutagenesis, the substitution near the N-terminus alters protein complex of frequency (FRQ) and white-collar complex (WCC)binding to FRH, but produces an unusual short clock period
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R806H
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interacts with the circadian oscillator component FREQUENCY (FRQ), but interaction between the FRQFRHR806H complex (FFC) and White Collar Complex is severely affected
E168A
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
E168Q
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
E168A
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site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
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site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
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site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
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site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
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E168A
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site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
-
E168Q
-
site-directed mutagenesis, the mutation eliminates ATP hydrolysis and helicase activity, and the mutant is unable to restore PexsD-lacZ activity to levels observed with wild-type DeaD
-
E909K
site-directed mutagenesis, the temperature-sensitive mutant, encoded by the slt22-1 allele, is synthetically lethal with mutations in U2 or U6 snRNAs that affect the stability or conformation of U2/U6 helix II. The ATPase activity of this variant is no longer stimulated by a U2/ U6 duplex, it is proposed that Brr2 might proofread U2/U6 interactions. The E909K exchange in Brr2 blocks splicing in extracts at or before the first catalytic step and leads to the appearance of an off-pathway spliceosomal particle following B complex formation, which lacks U4 and U5 snRNAs
G858R
site-directed mutagenesis, the mutant shows differing cross-linking profiles compared to wild-type Brr2, the mutation is in the NC 5'HP/separator loop with U6 snRNA
K177A
mutant enzyme shows no stimulation of ATPase activity by single-stranded RNA
R681C
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
R681H
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
V683L
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
Y689C
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the beginning of the RecA2 domain
E909K
-
site-directed mutagenesis, the temperature-sensitive mutant, encoded by the slt22-1 allele, is synthetically lethal with mutations in U2 or U6 snRNAs that affect the stability or conformation of U2/U6 helix II. The ATPase activity of this variant is no longer stimulated by a U2/ U6 duplex, it is proposed that Brr2 might proofread U2/U6 interactions. The E909K exchange in Brr2 blocks splicing in extracts at or before the first catalytic step and leads to the appearance of an off-pathway spliceosomal particle following B complex formation, which lacks U4 and U5 snRNAs
-
G858R
-
site-directed mutagenesis, the mutant shows differing cross-linking profiles compared to wild-type Brr2, the mutation is in the NC 5'HP/separator loop with U6 snRNA
-
R681C
-
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
-
R681H
-
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
-
V683L
-
site-directed mutagenesis, a brr2 mutation linked to the RP33 form of autosomal dominant retinitis pigmentosa, it maps to the linker between the RecA domains of the NC, the mutation leads to altered Brr2 ATPase activity and aberrant partitioning of spliceosomes along activation and discard pathways
-
E300A
-
mutation causes severe defect in RNA unwinding that correlates with reduced rate of ATP hydrolysis
H299A
-
mutation elicits defects in RNA unwinding but spares the ATPase activity
K191A
-
mutation causes severe defect in RNA unwinding that correlates with reduced rate of ATP hydrolysis
R229A
-
mutation causes severe defect in RNA unwinding that correlates with reduced rate of ATP hydrolysis
T192A
-
mutation causes severe defect in RNA unwinding that correlates with reduced rate of ATP hydrolysis
T326A
-
mutation elicits defects in RNA unwinding but spares the ATPase activity
T328A
-
mutation elicits defects in RNA unwinding but spares the ATPase activity
D172A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 41% of the ratio determined for the wild-type enzyme
E169A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 38% of the ratio determined for the wild-type enzyme
E173A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 17% of the ratio determined for the wild-type enzyme
E180A
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the ration of (kcat/Km)ATP/(kcat/Km)GTP is 35% of the ratio determined for the wild-type enzyme
E182A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 29% of the ratio determined for the wild-type enzyme
F179A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 19% of the ratio determined for the wild-type enzyme
K186A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 15% of the ratio determined for the wild-type enzyme
K187A
-
the ration of (kcat/Km)ATP/(kcat/Km)GTP is 8% of the ratio determined for the wild-type enzyme
Q188A
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the ration of (kcat/Km)ATP/(kcat/Km)GTP is 33% of the ratio determined for the wild-type enzyme
R170A
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the ration of (kcat/Km)ATP/(kcat/Km)GTP is 31% of the ratio determined for the wild-type enzyme
additional information
K232A
site-directed mutagenesis in the helicase domain of NS3
K199A/T200A
-
site-directed mutagenesis, mutant avoid of basal and of RNA-stimulated NTPase activity
K199A/T200A
-
site-directed mutagenesis, the mutation in the C-terminal domain of NS3 eliminates both the basal and the RNA-stimulated NTPase activity
K199A/T200A
-
site-directed mutagenesis, mutant avoid of basal and of RNA-stimulated NTPase activity
-
K199A/T200A
-
site-directed mutagenesis, the mutation in the C-terminal domain of NS3 eliminates both the basal and the RNA-stimulated NTPase activity
-
M1708A
NS3-4A construct, ability to bind and unwind RNA in vitro, mutation reduces functional NS3-4A binding affinity for RNA by 500-fold relative to the wild-type
M1708A
site-directed mutagenesis, the NS3-4A mutant shows decreased ATPase activity and reduced RNA stimulation activity compared to wild-type NS3
S1369R
NS3-4A construct, suppressor mutant, ATP-coupled RNA affinity identical to that of wild-type NS3-4A
S1369R
site-directed mutagenesis, the NS3-4A mutant shows increased ATPase activity and RNA stimulation activity compared to wild-type NS3
S1369R/M1708A
NS3-4A construct, reduced ATP-coupled RNA affinity of the single mutant suppressed by the addition of the S1369R mutation
S1369R/M1708A
site-directed mutagenesis, the NS3-4A mutant shows increased ATPase activity and reduced RNA stimulation activity compared to wild-type NS3
S1369R/Y1702A
NS3-4A construct, reduced ATP-coupled RNA affinity of the single mutant suppressed by the addition of the S1369R mutation
S1369R/Y1702A
site-directed mutagenesis, the NS3-4A mutant shows decreased ATPase activity and reduced RNA stimulation activity compared to wild-type NS3
Y1702A
NS3-4A construct, ability to bind and unwind RNA in vitro, mutation reduces functional NS3-4A binding affinity for RNA by 500-fold relative to the wild-type
Y1702A
site-directed mutagenesis, the NS3-4A mutant shows decreased ATPase activity and reduced RNA stimulation activity compared to wild-type NS3
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construction of double knockdown mutant rh3-4/clpr2-1, chloroplast rps12-int1 splicing defects in mutant rh3-4
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generation of RH7 knockout mutant lines, several morphological alterations such as disturbed vein pattern, pointed first true leaves, and short roots, which resemble ribosome-related mutants of Arabidopsis thaliana, phenotype analysis of rh7-5 and rh7-8 mutants under 22°C, overview. Knockout mutants of AtRH7 display several morphological alterations during vegetative and reproductive growth. In addition, the mutants exhibit severe defects in germination and leaf development under long-term low temperature conditions. Accumulation of rRNA precursors in rh7 mutant plants corroborate the hypothesis that AtRH7 affects ribosome biogenesis. AtRH7 mutations affect ribosomal RNA biogenesis in the nucleolus
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generation of RH7 knockout mutant lines, several morphological alterations such as disturbed vein pattern, pointed first true leaves, and short roots, which resemble ribosome-related mutants of Arabidopsis thaliana, phenotype analysis of rh7-5 and rh7-8 mutants under 22°C, overview. Knockout mutants of AtRH7 display several morphological alterations during vegetative and reproductive growth. In addition, the mutants exhibit severe defects in germination and leaf development under long-term low temperature conditions. Accumulation of rRNA precursors in rh7 mutant plants corroborate the hypothesis that AtRH7 affects ribosome biogenesis. AtRH7 mutations affect ribosomal RNA biogenesis in the nucleolus
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virus-induced ISE2 gene silencing, TRV constructs are introduced into Agrobacterium strain GV3101 for plant infiltrations. Loss of ISE2 compromises C-to-U RNA editing at specific sites. Total RNA is isolated from leaves of the same age as the ISE2-SUP leaves analyzed, and RNA editing at the six candidate ISE2-specific sites and several unaffected sites is measured. The clpP1-559 site exhibited reduced RNA editing efficiency in both the ISE2-SUP and the var2-2 leaves, but the extent of RNA editing in ISE2-SUP leaves (25%) is less than half of that in var2-2 leaves (59%). Thus, depletion of ISE2 has a more drastic effect on the RNA editing efficiency at the clpP1-559 site than depletion of FTSH2. Similarly, while the var2-2 mutant displays decreased RNA editing at petL-5 and rpoA-200, the decrease at those sites is more drastic in ISE2-SUP leaves than in var2-2 leaves. Accumulation of chloroplast mRNAs is altered in the absence of ISE2
additional information
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virus-induced ISE2 gene silencing, TRV constructs are introduced into Agrobacterium strain GV3101 for plant infiltrations. Loss of ISE2 compromises C-to-U RNA editing at specific sites. Total RNA is isolated from leaves of the same age as the ISE2-SUP leaves analyzed, and RNA editing at the six candidate ISE2-specific sites and several unaffected sites is measured. The clpP1-559 site exhibited reduced RNA editing efficiency in both the ISE2-SUP and the var2-2 leaves, but the extent of RNA editing in ISE2-SUP leaves (25%) is less than half of that in var2-2 leaves (59%). Thus, depletion of ISE2 has a more drastic effect on the RNA editing efficiency at the clpP1-559 site than depletion of FTSH2. Similarly, while the var2-2 mutant displays decreased RNA editing at petL-5 and rpoA-200, the decrease at those sites is more drastic in ISE2-SUP leaves than in var2-2 leaves. Accumulation of chloroplast mRNAs is altered in the absence of ISE2
additional information
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virus-induced ISE2 gene silencing, TRV constructs are introduced into Agrobacterium strain GV3101 for plant infiltrations. Loss of ISE2 compromises C-to-U RNA editing at specific sites. Total RNA is isolated from leaves of the same age as the ISE2-SUP leaves analyzed, and RNA editing at the six candidate ISE2-specific sites and several unaffected sites is measured. The clpP1-559 site exhibited reduced RNA editing efficiency in both the ISE2-SUP and the var2-2 leaves, but the extent of RNA editing in ISE2-SUP leaves (25%) is less than half of that in var2-2 leaves (59%). Thus, depletion of ISE2 has a more drastic effect on the RNA editing efficiency at the clpP1-559 site than depletion of FTSH2. Similarly, while the var2-2 mutant displays decreased RNA editing at petL-5 and rpoA-200, the decrease at those sites is more drastic in ISE2-SUP leaves than in var2-2 leaves. Accumulation of chloroplast mRNAs is altered in the absence of ISE2
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trans interference by BMV 1a protein helicase mutants with BMV 1a protein-stimulated RNA3 accumulation, overview
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knockdown of hel-1 specifically decreases daf-2(-)-mediated longevity in an RNAi-hypersensitive rrf-3(pk1426) mutant background. hel-1 RNAi has a robust and specific effect on the longevity of daf-2 mutants. Resistance to heat stress is not affected by hel-1 mutations or RNAi, effects of hel-1 RNAi and hel-1 mutations on resistance to pathogenic bacteria (Pseudomonas aeruginosa, PA14) and oxidative stress are variable
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knockdown of hel-1 specifically decreases daf-2(-)-mediated longevity in an RNAi-hypersensitive rrf-3(pk1426) mutant background. hel-1 RNAi has a robust and specific effect on the longevity of daf-2 mutants. Resistance to heat stress is not affected by hel-1 mutations or RNAi, effects of hel-1 RNAi and hel-1 mutations on resistance to pathogenic bacteria (Pseudomonas aeruginosa, PA14) and oxidative stress are variable
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for the truncated NS3 helicase domain both NTPase and helicase activities are up-regulated by NS5B, for the full-length NS3, the NTPase activity, but not the helicase activity, is stimulated by NS5B, specific interaction between NS3 and NS5B
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for the truncated NS3 helicase domain both NTPase and helicase activities are up-regulated by NS5B, for the full-length NS3, the NTPase activity, but not the helicase activity, is stimulated by NS5B, specific interaction between NS3 and NS5B
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for the truncated NS3 helicase domain both NTPase and helicase activities are up-regulated by NS5B, for the full-length NS3, the NTPase activity, but not the helicase activity, is stimulated by NS5B, specific interaction between NS3 and NS5B
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ddx27 mutant zebrafish show skeletal muscle abnormalities. Overexpression of human DDX27 mRNA results in a significant decrease in mutant zebrafish phenotype
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depletion of the enzyme gene expression by RNAi in both glia and clock neurons, knockdown of belle with the pan-glial driver repo-Gal4 and lama-Gal4, a driver for glial precursor cells, lamina precursor cells and lamina neurons. The belEY08943 mutant exhibits a less striking phenotype: a significant difference was observed in the PDF positive s-LNvs only in constant darkness, where the oscillation of PER is 4 h delayed compared to control, similarly to what described in belcap-1 flies. Both belle mutants are characterized by a reduction in the number of l-LNvs: a high percentage of brains, in fact, presented three neurons instead of the canonical four (83.93, 76.28, and 6.78% in bel cap-1, bel EY08943, and white 1118, respectively). In contrast, RNAi against belle in the photoreceptor cells (GMR-Gal4 and ninaE-Gal4) does not affect either vitality or behavior. Locomotor activity of belle knockdown flies under different conditions, overview
additional information
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depletion of the enzyme gene expression by RNAi in both glia and clock neurons, knockdown of belle with the pan-glial driver repo-Gal4 and lama-Gal4, a driver for glial precursor cells, lamina precursor cells and lamina neurons. The belEY08943 mutant exhibits a less striking phenotype: a significant difference was observed in the PDF positive s-LNvs only in constant darkness, where the oscillation of PER is 4 h delayed compared to control, similarly to what described in belcap-1 flies. Both belle mutants are characterized by a reduction in the number of l-LNvs: a high percentage of brains, in fact, presented three neurons instead of the canonical four (83.93, 76.28, and 6.78% in bel cap-1, bel EY08943, and white 1118, respectively). In contrast, RNAi against belle in the photoreceptor cells (GMR-Gal4 and ninaE-Gal4) does not affect either vitality or behavior. Locomotor activity of belle knockdown flies under different conditions, overview
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in order to increase the peptide linker region length, a 23 amino acid residue polypeptide with a composition of NASSGSSASSPSASNSPGANGSS was inserted between the native interdomain region's Ala and Thr residues. The sequence of the new extended interdomain linker is PANSSIANASSGSSASSPSASNSPGANGSSTLEAE. This peptide sequence is chosen because it has similar structural and dynamic properties to the native interdomain region, and both the designed and the native interdomain linker are predicted to form a flexible and unstructured region. In addition, since DbpA is purified as a native protein, small and polar amino acids, which promote peptide solubility, are placed into the polypeptide insert to discourage the aggregation of extended DbpA and its partition into inclusion bodies. The new interdomain linker is not digested by the Escherichia coli proteolytic enzymes and the extended DbpA is expressed as an intact and soluble protein. Breaking the sequence of the interdomain peptide linker and inserting the 23 amino acids peptide segment causes a decrease in binding affinity, likely as a consequence of formation of non-native interaction between the insert peptide and the RNA molecule or other regions of the protein and not a consequence of disrupting native interactions between the DbpA RNA binding domain and the interdomain linker. The peptide extension is not effecting the formation of the proper ATP pocket, but the ATP turnover rate is affected by the peptide extension. Although the ATP turnover of the extended DbpA is reduced when compared to wild-type DbpA, extended DbpA is a much more efficient enzyme than many members of DEAD-box family of proteins. The reduction on the ATP turnover of the extended DbpA is a consequence of its decrease in binding affinity for RNA. The extension of the interdomain linker region has no effect on the ability of DbpA to perform its helicase function. Thus, the physical connection of DbpA RNA binding domain to the catalytic core is unimportant for the helicase activity of DbpA, suggesting the DbpA protein is a region-specific enzyme, which would unwind any double-helix substrate near hairpin 92
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construction of the NS3-4A mutant affected in its acidic domain, the mutant shows altered RNA binding and increased ATPase activity, kinetics, overview
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construction of the NS3-4A mutant affected in its acidic domain, the mutant shows altered RNA binding and increased ATPase activity, kinetics, overview
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mutagenesis of conserved p54 helicase motifs activates translation in the tethered function assay, reduces accumulation of p54 in P-bodies in HeLa cells, and inhibits its capacity to assemble P-bodies in p54-depleted cells
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mutagenesis of conserved p54 helicase motifs activates translation in the tethered function assay, reduces accumulation of p54 in P-bodies in HeLa cells, and inhibits its capacity to assemble P-bodies in p54-depleted cells
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recombinant RHA domains are evaluated for binding activity to post-transcriptional control element (PCE) in comparison with nonfunctional PCE and generic control double-stranded RNAs (dsRNAs). N-terminal domain exhibits higher binding affinity for PCE than for nonfunctional mutant RNA or control dsRNA. Highly conserved surface-exposed lysine residues are required for selective interaction with PCE RNA. In cells, the N-terminal domain directs interaction with PCE, mRNA and its exogenous expression blocks the translation activity of endogenous RHA
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wild-type and mutant enzymes in vitro RNA binding and unwinding or in the cell during HIV-1 production during RNA helicase A-RNA interaction and RNA helicase A-stimulated viral RNA processes, overview. The mutations do not prevent RNA helicase A from binding to HIV-1 RNA in vitro aswell, but dramatically reduce RNA helicase A-HIV-1 RNA interaction in the cells
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generation of codon-optimized DNA fragments encoding selected regions of hBrr2
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generation of the DELTAUPF1_1x02loop and UPF1_1DELTACHx02loop mutants, and of truncation mutant UPF1_2DELTACH
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isolation of the catalytically active helicase core DHX8DELTA547 enzyme fragment. The structure of DHX8DELTA547 reveals flexibility in the DEAH motif. The two DHX8DELTA547-ADP structures reveal a domain organisation very similar to that of other DEAH-box helicases. DHX8DELTA547 adopts a pyramidal-like structure with the two N-terminal RecA domains and the C-terminal ratchet-like and OB-fold domains on opposite sides of the putative DHX8 RNA-binding tunnel, and the C- and N-terminal domains connected by the WH domain. Comparison of kinetics of wild-type full-length enzyme and truncated mutant, overview
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isolation of the catalytically active helicase core DHX8DELTA547 enzyme fragment. The structure of DHX8DELTA547 reveals flexibility in the DEAH motif. The two DHX8DELTA547-ADP structures reveal a domain organisation very similar to that of other DEAH-box helicases. DHX8DELTA547 adopts a pyramidal-like structure with the two N-terminal RecA domains and the C-terminal ratchet-like and OB-fold domains on opposite sides of the putative DHX8 RNA-binding tunnel, and the C- and N-terminal domains connected by the WH domain. Comparison of kinetics of wild-type full-length enzyme and truncated mutant, overview
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knockout of enzyme Lmo1722 decreases the number of mature 70S ribosomes at low temperatures and decreases Listeria monocytogenes motility by downregulating flaA expression
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knockout of enzyme Lmo1722 decreases the number of mature 70S ribosomes at low temperatures and decreases Listeria monocytogenes motility by downregulating flaA expression
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knockout of enzyme Lmo1722 decreases the number of mature 70S ribosomes at low temperatures and decreases Listeria monocytogenes motility by downregulating flaA expression
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trageting of the enzyme DDX6 with shRNA in N2a cells. Construction of a mutant DDX6 that contains a point mutation within the DEAD box motif II of DDX6 and thus lacks helicase activity, but is still able to coimmunoprecipitate TRIM32
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generation of Ddx3xfl/fl mice, analysis of DDX3X and DDX3Y activity in fibroblasts from gene-targeted mice. Mouse embryonic fibroblasts (MEFs) derived from female Ddx3xfl/fl CreERT2 mice are treated with 4-OHT to delete Ddx3x. Innate immunity of DDX3X-deficient cells and of Ddx3xfl/y Vav-iCre mice is analyzed, overview. Mice lacking DDX3X in hematopoietic cells have reduced numbers of lymphocytes and natural killer cells. Mice lacking DDX3X in the hematopoietic system produce reduced amounts of serum IL-12 and IFNgamma after Listeria monocytogenes infection compromising the immune response of macrophages
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plasmids and siRNAs are transfected in HEK-293T cells using calcium phosphate protocol. NIH 3T3 mouse fibroblasts, U2OS, and murine embryonic stem cells are transfected with the siRNA, siRNA-mediated depletions and synthetic RNA levels are monitored by quantitative RT-PCR expression analysis 3-4 days post transfection
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114)
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the N-terminal part of the TGBp1 NTPase/helicase domain comprising conserved motifs I, Ia and II is sufficient for ATP hydrolysis, RNA binding and homologous proteinprotein interactions. Point mutations in a single conserved basic amino acid residue upstream of motif I have little effect on the activities of C-terminally truncated mutants of both TGBp1 proteins. When introduced into the full-length NTPase/helicase domains, these mutations cause a substantial decrease in the ATPase activity of the protein, suggesting that the conserved basic amino acid residue upstream of motif I is required to maintain a reaction-competent conformation of the TGBp1 ATPase active site
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the N-terminal part of the TGBp1 NTPase/helicase domain comprising conserved motifs I, Ia and II is sufficient for ATP hydrolysis, RNA binding and homologous proteinprotein interactions. Point mutations in a single conserved basic amino acid residue upstream of motif I have little effect on the activities of C-terminally truncated mutants of both TGBp1 proteins. When introduced into the full-length NTPase/helicase domains, these mutations cause a substantial decrease in the ATPase activity of the protein, suggesting that the conserved basic amino acid residue upstream of motif I is required to maintain a reaction-competent conformation of the TGBp1 ATPase active site
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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construction of an deletion mutant DELTAdeaD
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an NS3 mutant, that is deficient in RNA binding and its associated RSS activity, is inactive in complementing the RNA silencing suppressor function of the Tat protein of Human immunodeficiency virus type 1
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an NS3 mutant, that is deficient in RNA binding and its associated RSS activity, is inactive in complementing the RNA silencing suppressor function of the Tat protein of Human immunodeficiency virus type 1
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an NS3 mutant, that is deficient in RNA binding and its associated RSS activity, is inactive in complementing the RNA silencing suppressor function of the Tat protein of Human immunodeficiency virus type 1
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construction of PCR-generated truncated fragments of cshA, including the ATPase and the helicase domains and ending in a stop codon. Genes more abundant in the wild-type than in the cshA mutant upon MazFsa expression, overview. Mutation of cshA affects growth and cell viability
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construction of PCR-generated truncated fragments of cshA, including the ATPase and the helicase domains and ending in a stop codon. Genes more abundant in the wild-type than in the cshA mutant upon MazFsa expression, overview. Mutation of cshA affects growth and cell viability
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construction of PCR-generated truncated fragments of cshA, including the ATPase and the helicase domains and ending in a stop codon. Genes more abundant in the wild-type than in the cshA mutant upon MazFsa expression, overview. Mutation of cshA affects growth and cell viability
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construction of PCR-generated truncated fragments of cshA, including the ATPase and the helicase domains and ending in a stop codon. Genes more abundant in the wild-type than in the cshA mutant upon MazFsa expression, overview. Mutation of cshA affects growth and cell viability
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additional information
Thermochaetoides thermophila
design of a mutant of Prp43 which allows us to trap the closed conformation by the introduction of an internal disulfide bond (ctPrp43-IDSB). For this purpose, one cysteine is introduced into the RecA1 domain and another one into the ratchet-like domain at exposed positions to maximize the number of formed disulfide bonds. The wild-type protein contains nine cysteines, the ctPrp43-IDSB mutant two additional ones, the majority of ctPrp43-IDSB exhibits the internal disulfide bridge. Prp43 trapped in the closed conformation is impaired in its helicase activity. The intrinsic ATPase activity of ctPrp43-IDSB is similar to the one determined for wild-type ctPrp43. ctPrp43-IDSB is also stimulated by ctPfa1-GP and by U16-RNA in the presence of the ctPfa1-GP, but in the contrast to wild-type Prp43 also just by U16-RNA. Prp43 in the trapped closed conformation appears to be more prone for the stimulation of the ATPase. Conformational rearrangements at the helicase core, structure analysis, overview
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Thermochaetoides thermophila CBS 144.50
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design of a mutant of Prp43 which allows us to trap the closed conformation by the introduction of an internal disulfide bond (ctPrp43-IDSB). For this purpose, one cysteine is introduced into the RecA1 domain and another one into the ratchet-like domain at exposed positions to maximize the number of formed disulfide bonds. The wild-type protein contains nine cysteines, the ctPrp43-IDSB mutant two additional ones, the majority of ctPrp43-IDSB exhibits the internal disulfide bridge. Prp43 trapped in the closed conformation is impaired in its helicase activity. The intrinsic ATPase activity of ctPrp43-IDSB is similar to the one determined for wild-type ctPrp43. ctPrp43-IDSB is also stimulated by ctPfa1-GP and by U16-RNA in the presence of the ctPfa1-GP, but in the contrast to wild-type Prp43 also just by U16-RNA. Prp43 in the trapped closed conformation appears to be more prone for the stimulation of the ATPase. Conformational rearrangements at the helicase core, structure analysis, overview
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Thermochaetoides thermophila DSM 1495
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design of a mutant of Prp43 which allows us to trap the closed conformation by the introduction of an internal disulfide bond (ctPrp43-IDSB). For this purpose, one cysteine is introduced into the RecA1 domain and another one into the ratchet-like domain at exposed positions to maximize the number of formed disulfide bonds. The wild-type protein contains nine cysteines, the ctPrp43-IDSB mutant two additional ones, the majority of ctPrp43-IDSB exhibits the internal disulfide bridge. Prp43 trapped in the closed conformation is impaired in its helicase activity. The intrinsic ATPase activity of ctPrp43-IDSB is similar to the one determined for wild-type ctPrp43. ctPrp43-IDSB is also stimulated by ctPfa1-GP and by U16-RNA in the presence of the ctPfa1-GP, but in the contrast to wild-type Prp43 also just by U16-RNA. Prp43 in the trapped closed conformation appears to be more prone for the stimulation of the ATPase. Conformational rearrangements at the helicase core, structure analysis, overview
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Thermochaetoides thermophila IMI 039719
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design of a mutant of Prp43 which allows us to trap the closed conformation by the introduction of an internal disulfide bond (ctPrp43-IDSB). For this purpose, one cysteine is introduced into the RecA1 domain and another one into the ratchet-like domain at exposed positions to maximize the number of formed disulfide bonds. The wild-type protein contains nine cysteines, the ctPrp43-IDSB mutant two additional ones, the majority of ctPrp43-IDSB exhibits the internal disulfide bridge. Prp43 trapped in the closed conformation is impaired in its helicase activity. The intrinsic ATPase activity of ctPrp43-IDSB is similar to the one determined for wild-type ctPrp43. ctPrp43-IDSB is also stimulated by ctPfa1-GP and by U16-RNA in the presence of the ctPfa1-GP, but in the contrast to wild-type Prp43 also just by U16-RNA. Prp43 in the trapped closed conformation appears to be more prone for the stimulation of the ATPase. Conformational rearrangements at the helicase core, structure analysis, overview
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