Information on EC 3.1.26.12 - ribonuclease E

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
3.1.26.12
-
RECOMMENDED NAME
GeneOntology No.
ribonuclease E
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
endonucleolytic cleavage of single-stranded RNA in A- and U-rich regions
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
tRNA processing
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
76106-82-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
; gene NCgl2281
-
-
Manually annotated by BRENDA team
gene NCgl2281
-
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Manually annotated by BRENDA team
strain 18-11BP, gene ams/rne/hmp1
-
-
Manually annotated by BRENDA team
strain BRL2288
-
-
Manually annotated by BRENDA team
strain CF881, gene rne
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Manually annotated by BRENDA team
strain GM402 BL21(DE3)
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Manually annotated by BRENDA team
strain JM109, gene rne
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Manually annotated by BRENDA team
strain CA244, gene rne
-
-
Manually annotated by BRENDA team
strain KSL2000
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-
Manually annotated by BRENDA team
strain MG1693 or derivatives, gene rne
UniProt
Manually annotated by BRENDA team
strain N3433
-
-
Manually annotated by BRENDA team
gene rne
Uniprot
Manually annotated by BRENDA team
strain ATCC 10004
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-
Manually annotated by BRENDA team
strain 8401/pRL1JI
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-
Manually annotated by BRENDA team
strain 3764, DSM 938
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-
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
13mer nucleotide sequence of RNAI + H2O
?
show the reaction diagram
-
endonucleolytic cleavage, a synthetic 13-nt oligoribonucleotide, representing the cleavage site of RNAI, from the 5' end, with the canonical RNase E cleavage site located between U5 and A6
-
-
?
16s rRNA + H2O
?
show the reaction diagram
23S rRNA + H2O
5.8S-like rRNA + ?
show the reaction diagram
23S rRNA + H2O
?
show the reaction diagram
5' monophosphorylated RNA oligonucleotides + H2O
?
show the reaction diagram
-
several synthetic substrates, overview
-
-
?
5'-capped RNA I.26 + H2O
?
show the reaction diagram
-
low activity, cleavage of the 5' substrate end
-
-
?
5'-GAGACAGUAUUUG + H2O
5'-GAGACAGU + AUUUG
show the reaction diagram
-
LU13 substrate, LU13 is a BR13 derivative that has the central G of the 5' triplet replaced with an A. 5'-biotinylated LU13 is cleaved more rapidly when conjugated to streptavidin prior to incubation with N-terminal half-RNase E. In the absence of streptavidin conjugation, 5'-biotinylated LU13 is cleaved as poorly as its 5' hydroxylated equivalent
-
-
?
5'-GGGACAGUAUUUG + H2O
5'-GGGACAGU + AUUUG
show the reaction diagram
-
BR13 substrate, RNase E can cleave certain RNAs rapidly without requiring a 5'-monophosphorylated end. Cleavage of 5'-hydroxylated oligonucleotide substrate by the N-terminal half of RNase E. RNase E can bind with higher affinity to a 5'-hydroxylated substrate with multiple single-stranded regions than to a 5'-monophosphorylated substrate with one single-stranded site
-
-
?
5'-GGGACAGUAUUUG-3' + H2O
?
show the reaction diagram
-
-
-
-
?
5'-labeled RNA oligonucleotides + H2O
?
show the reaction diagram
-
synthetic RNA substrate variants based on known enzyme RNA substrate sequences, recombinant Rne498 catalytic domain, cleavage site specificity, overview
-
-
?
5'-triphosphorylated cspA mRNA + H2O
?
show the reaction diagram
-
RNase E recognizes multiple single-stranded regions in cspA mRNA
-
-
?
5'-triphosphorylated epd-pgk RNA + H2O
?
show the reaction diagram
-
RNase E cleavage of 5'-triphosphorylated epd-pgkRNA is faster than 5'-triphosphorylated 9S RNA and RNAi, but not as fast as the rate of cleavage of 5'-triphosphorylated cspA mRNA
-
-
?
5'-triphosphorylated mRNA fragment + H2O
?
show the reaction diagram
-
-
-
-
?
5'-triphosphorylated RNAi + H2O
?
show the reaction diagram
-
-
-
-
?
5'ACAGUAUUUG-fluorescein + H2O
5'ACAGU + AUUUG-fluorescein
show the reaction diagram
-
5' monophosphorylated, 3' fluorescein-labeled synthetic substrate with protective 2'-O-methyl groups at all positions based on the 5' cleavage site of pBR322 RNA I
-
-
?
5'AUCAAAGAAA + H2O
5'AUCAAAGA + AA
show the reaction diagram
-
5'-labeled synthetic RNA substrate, modified 9S RNA sequence, recombinant Rne498 catalytic domain, no activity with the wild-type 9S RNA sequence 5'AUCAAAUAAA and with modified sequence 5'AUCAGAUAAA
-
-
?
5'AUCAAGUAAA + H2O
5'AUCAAGU + AAA
show the reaction diagram
-
low activity, 5'-labeled synthetic RNA substrate, modified 9S RNA sequence, recombinant Rne498 catalytic domain, no activity with the wild-type 9S RNA sequence 5'AUCAAAUAAA and with modified sequence 5'AUCAGAUAAA
-
-
?
5'AUCGAAUAAA + H2O
5'AUCGA + AUAAA
show the reaction diagram
-
5'-labeled synthetic RNA substrate, modified 9S RNA sequence, recombinant Rne498 catalytic domain, no activity with the wild-type 9S RNA sequence 5'AUCAAAUAAA and with modified sequence 5'AUCAGAUAAA
-
-
?
5'GGGA(D-dT)CAGUAUUU-fluorescein + H2O
?
show the reaction diagram
-
5' monophosphorylated, 3' fluorescein-labeled synthetic substrate with protective 2'-O-methyl groups at all positions based on the 5' cleavage site of RNA I
-
-
?
82 nt of the NifA mRNA + H2O
?
show the reaction diagram
9S mRNA + H2O
p5S RNA + ?
show the reaction diagram
-
-
-
-
?
9S precursor RNA + H2O
p5S RNA + ?
show the reaction diagram
-
-
-
-
?
9S RNA + H2O
5S RNA + ?
show the reaction diagram
-
-
-
-
?
9S RNA + H2O
?
show the reaction diagram
9S rRNA + H2O
?
show the reaction diagram
9S rRNA precursor + H2O
5S rRNA + ?
show the reaction diagram
9S-RNA + H2O
?
show the reaction diagram
9SA RNA + H2O
?
show the reaction diagram
AAUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
Aquifex aeolicus 9S rRNA + H2O
?
show the reaction diagram
-
-
-
-
?
Bacillus subtilis aprE leader-lacZ mRNA + H2O
?
show the reaction diagram
-
wild-type and mutant substrate, the latter with an exchange of a G and an A at +31 and +32, respectively, cleavage of the Bacillus subtilis transcript in a structure-dependent manner at the 5' end to the U residue at +12 within a double-stranded segment of an AU-rich sequence, which is part of the stem-loop structure at the 5' end of the transcript
-
-
?
bacteriophage T4 gene 32 mRNA + H2O
?
show the reaction diagram
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processing at the -71 site, which forms a stem-loop essential for enzyme activity of RNase E, the putative consensus sequence is RAUUW, mutational disruption of the stem-loop leads to loss of activity, mechanism, overview
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-
?
BR13 + H2O
?
show the reaction diagram
BR13 RNA + H2O
?
show the reaction diagram
BR13N RNA + H2O
?
show the reaction diagram
-
synthetic RNA substrate, the cleavage site is GGGACAGUCUGUG
-
-
?
BR30M + H2O
?
show the reaction diagram
CAUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
cspA mRNA + H2O
?
show the reaction diagram
endonulceolytic cleavage of sodB mRNA
?
show the reaction diagram
-
the enzyme cleaves within the sodB 5'-untranslated region in vitro, thereby removing the 5' stem-loop structure that facilitates Hfq and ribosome binding, RNase E cleavage can also occur at a cryptic site that becomes available upon sodB 5'-UTR/RyhB base pairing
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-
?
Escherichia coli M1 RNA + H2O
?
show the reaction diagram
-
-
-
-
?
fluorogenic oligonucleotides + H2O
?
show the reaction diagram
-
5' monophosphorylated or 5' hydroxylated substrates, P-BR14-FD or OH-BR14-FD
-
-
?
GAUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
GUUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
immature 16S rRNA + H2O
mature 16S rRNA
show the reaction diagram
MicX + H2O
?
show the reaction diagram
-
endonucleolytic cleavage, wild-type substrate, RNase E-dependent processing stabilizes MicX, a Vibrio cholerae sRNA
-
-
?
MicX sRNA + H2O
?
show the reaction diagram
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cleavage involves protein Hfq
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?
MicX_DELTA196-263 mutant + H2O
?
show the reaction diagram
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endonucleolytic cleavage, a truncated Vibrio cholerae sRNA
-
-
?
mRNA + H2O
?
show the reaction diagram
-
-
-
-
?
Omp11 RNA + H2O
?
show the reaction diagram
-
-
-
-
?
ompA mRNA + H2O
?
show the reaction diagram
p23 RNA + H2O
?
show the reaction diagram
pppRNA I.26 + H2O
?
show the reaction diagram
-
low activity, cleavage of the 5' substrate end
-
-
?
pre-5S rRNA + H2O
?
show the reaction diagram
pre-tRNACys + H2O
tRNACys + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pre-tRNAHis + H2O
tRNAHis + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pre-tRNAPro + H2O
tRNAPro + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pRNA I.26 + H2O
?
show the reaction diagram
-
a monophosphate at the 5' end of the RNA I substrate stimulates the enzyme 25-30fold, cleavage of the 5' substrate end
-
-
?
pSu3 + H2O
?
show the reaction diagram
puf mRNA + H2O
?
show the reaction diagram
Rep mRNA + H2O
?
show the reaction diagram
-
the arginine-rich RNA binding domain and the protein scaffold domain of RNase E are dispensable for degradation of the replication initiator protein (Rep) mRNA of the ColE2 plasmid
-
-
?
RNA + H2O
?
show the reaction diagram
RNA I + H2O
?
show the reaction diagram
RNA I.26 + H2O
?
show the reaction diagram
rne mRNA + H2O
?
show the reaction diagram
-
the enzyme autoregulates its expression by cleavage and processing of its own rne mRNA
-
-
?
rne-lacZ transcript + H2O
?
show the reaction diagram
-
-
-
-
?
rpsT mRNA + H2O
?
show the reaction diagram
S20 mRNA + H2O
?
show the reaction diagram
-
mRNA encoding ribosomal proteins, a single cleavage site at residues 300/301 is preceded by variable 5' extensions
-
-
?
S20 mRNA t160D + H2O
?
show the reaction diagram
-
ribosomal protein encoding RNA
-
-
?
S20 mRNA t175D + H2O
?
show the reaction diagram
-
ribosomal protein encoding RNA, structure mapping, secondary structure modeling, overview
-
-
?
S20 mRNA t194D + H2O
?
show the reaction diagram
-
ribosomal protein encoding RNA, structure mapping, secondary structure modeling, overview
-
-
?
S20 mRNA t84D + H2O
?
show the reaction diagram
-
ribosomal protein encoding RNA, contains a 5' stem loop with three noncanonical A-G pairs, structure mapping, secondary structure modeling, overview
-
-
?
S20 mRNA t87D + H2O
?
show the reaction diagram
S20 mRNA t95D + H2O
?
show the reaction diagram
-
processing
-
-
?
single-stranded RNA + H2O
?
show the reaction diagram
sodB mRNA + H2O
?
show the reaction diagram
-
RNase E and RNase III are required for sodB RNA decay in vivo
-
-
?
sodB192 mRNA + H2O
?
show the reaction diagram
-
cleavage of the 5'-untranslated region in vitro thereby removing the stem loop structure that facilitates Hfq and ribosome binding, additional cleavage at a cryptic site, that becomes available upon sodB5'-UTR/RyhB base pairing, RyhB is a small regulatory RNA involved in sodB translation control, overview
-
-
?
sRNA MicX_2-346 + H2O
?
show the reaction diagram
-
a truncated Vibrio cholerae sRNA
-
-
?
tRNA precursors + H2O
?
show the reaction diagram
-
the enzyme, cuts intercistronic regions of putative tRNA precursors, overview
-
-
?
tRNAArgHisLeuPro precursor + H2O
tRNAArg + tRNAHis + tRNALeu + tRNAPro + ?
show the reaction diagram
tRNAAsn precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAGlyCysLeu precursor + H2O
tRNAGly + tRNACys + tRNALeu + ?
show the reaction diagram
tRNAHis precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAPhe precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAPro precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNATyr precursor + H2O
tRNATyr + ?
show the reaction diagram
tRNATyrSu3 precursor + H2O
?
show the reaction diagram
tRNATyrsu3+ + H2O
tRNATyrsu3+ + ?
show the reaction diagram
-
a construct of 404 nucleotides containing a leader sequence and the amber suppressor form of tRNATyr
-
-
?
UAUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
-
G378 mutant substrate, p23 RNA variant derived from linearized DraI plasmid, in vitro substrate synthesis by SP6 RNA poylmerase
-
-
?
unc mRNA + H2O
?
show the reaction diagram
upRNA + H2O
?
show the reaction diagram
UUUUU-containing RNA oligonucleotide + H2O
?
show the reaction diagram
-
G378/A379 mutant substrate, p23 RNA variant derived from linearized DraI DN1 or DN34 plasmids, in vitro substrate synthesis by SP6 RNA poylmerase
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
23S rRNA + H2O
?
show the reaction diagram
9S rRNA + H2O
?
show the reaction diagram
-
the enzyme is essential for 9S RNA processing
-
-
?
9S rRNA precursor + H2O
5S rRNA + ?
show the reaction diagram
9S-RNA + H2O
?
show the reaction diagram
Aquifex aeolicus 9S rRNA + H2O
?
show the reaction diagram
-
-
-
-
?
Bacillus subtilis aprE leader-lacZ mRNA + H2O
?
show the reaction diagram
-
wild-type and mutant substrate, the latter with an exchange of a G and an A at +31 and +32, respectively, cleavage of the Bacillus subtilis transcript in a structure-dependent manner at the 5' end to the U residue at +12 within a double-stranded segment of an AU-rich sequence, which is part of the stem-loop structure at the 5' end of the transcript
-
-
?
BR13 + H2O
?
show the reaction diagram
-
endonucleolytic cleavage, BR13 is an oligoribonucleotide that contains the RNase E-cleaved sequence of RNA I
-
-
?
cspA mRNA + H2O
?
show the reaction diagram
-
degradation of the cspA mRNA in vivo is very rapid at temperatures greater than 30°C, overview
-
-
?
Escherichia coli M1 RNA + H2O
?
show the reaction diagram
-
-
-
-
?
immature 16S rRNA + H2O
mature 16S rRNA
show the reaction diagram
-
RNase G, i.e. CafA protein, and RNase E are both required for the 5' maturation of 16S ribosomal RNA
-
-
?
MicX sRNA + H2O
?
show the reaction diagram
-
cleavage involves protein Hfq
-
-
?
ompA mRNA + H2O
?
show the reaction diagram
-
ompA at a site which is rate determining for degradation and also cleaved by RNase K
-
-
?
p23 RNA + H2O
?
show the reaction diagram
-
endonucleolytic cleavage
-
-
?
pre-5S rRNA + H2O
?
show the reaction diagram
pre-tRNACys + H2O
tRNACys + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pre-tRNAHis + H2O
tRNAHis + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pre-tRNAPro + H2O
tRNAPro + 3'-leader of tRNA
show the reaction diagram
-
-
-
-
?
pSu3 + H2O
?
show the reaction diagram
-
endonucleolytic cleavage, the precursor of the Escherichia coli tRNATyrSu3, cleavage upstream of the RNase P cleavage site in vitro and in vivo
-
-
?
puf mRNA + H2O
?
show the reaction diagram
RNA + H2O
?
show the reaction diagram
rne mRNA + H2O
?
show the reaction diagram
-
the enzyme autoregulates its expression by cleavage and processing of its own rne mRNA
-
-
?
rpsT mRNA + H2O
?
show the reaction diagram
single-stranded RNA + H2O
?
show the reaction diagram
-
RhlB is an ATP-dependent motor that unfolds structured RNA for destruction by partner ribonucleases, RhlB associates with the essential endoribonuclease RNase E as part of the multi-enzyme RNA degradosome assembly, RNase E activates RhlB severalfold, determination and analysis of the specific protein interaction sites using limited protease digestion, domain cross-linking and homology modelling. The stoichiometry for RhlB-CTD/RNase E, residues 628-843, complex is 1:1, overview
-
-
?
sodB mRNA + H2O
?
show the reaction diagram
-
RNase E and RNase III are required for sodB RNA decay in vivo
-
-
?
tRNAArgHisLeuPro precursor + H2O
tRNAArg + tRNAHis + tRNALeu + tRNAPro + ?
show the reaction diagram
tRNAAsn precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAGlyCysLeu precursor + H2O
tRNAGly + tRNACys + tRNALeu + ?
show the reaction diagram
tRNAHis precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAPhe precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNAPro precursor + H2O
?
show the reaction diagram
-
-
-
-
?
tRNATyr precursor + H2O
tRNATyr + ?
show the reaction diagram
-
maturation, cleavage of the tyrT transcript, containing two tRNATyr1 sequences separated by a 209-nt spacer region plus a downstream mRNA, at three sites in the speacer region, overview
-
-
?
tRNATyrSu3 precursor + H2O
?
show the reaction diagram
-
cleavage in the 5' leader sequence, the enzyme is involved in regulation of cellular tRNA levels
-
-
?
unc mRNA + H2O
?
show the reaction diagram
-
the unc operon encodes the eight subunits of the Escherichia coli F1F0-ATPase, processing of the unc mRNAs by the RNase E, overview, RNase E is essential for uncC processing
-
-
?
upRNA + H2O
?
show the reaction diagram
-
RNase E is a processing enzyme involved in 3' end formation of M1 RNA, and plays a dual role in processing and degradation to achieve tight control of M1 RNA biosynthesis
-
-
-
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
100 mM, required
Mn2+
-
enzyme assay in presence of 1 mM MnCl2
additional information
-
enzyme assay in presence of Mg2+ and NH4Cl
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Diamide
-
treatment of the N-terminal catalytic domain with diamide causes complete loss of the zinc, but only slightly reduced activity as tetramer
EDTA
-
disrupts the oligomer
ribosomal protein L4
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L4 interacts with a site outside of the catalytic domain at the C-terminal domain of RNase E to regulate the endoribonucleolytic functions of the enzyme thus inhibiting RNase E-specific cleavage in vitro
-
RraA
-
RraAV1
-
the coexpression of RraAV1 more efficiently inhibits RNase E action than coexpression of RraA
-
RraAV2
-
RNA substrate-dependent inhibition of RraAV2 on RNase E
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RraB
-
in vitro cleavage of p23 RNA by 70 ng RNAse ES is inhibited by 20.3% by RraB
-
structured RNA
-
inhibits the plant enzyme
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
chaperone Hfq
-
plays an important role in facilitating the interaction ofRyhB with sodB mRNA, Hfq is not tightly retained by the RyhB-sodB mRNA complex and can be released from it through interaction with other RNAs added in trans
-
Hfq
-
a factor binding to the sodB RNA and facilitating cleavage due to induced conformational changes within the 5'-UTR, overview
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00033
5' hydroxylated fluorogenic oligonucleotide
-
pH 7.5, 25°C, recombinant N-terminal domain
-
0.00023
5' monophosphorylated fluorogenic oligonucleotide
-
pH 7.5, 25°C, recombinant N-terminal domain
-
0.008
5'-GAGACAGUAUUUG
-
5'-hydroxylated LU13
0.00013 - 0.0036
5'-GGGACAGUAUUUG
0.00207
5'-GGGACAGUAUUUG-3'
-
in 25 mM bis-Tris-propane (pH 8.3), 15 mM MgCl2, 100 mM NaCl, 0.1% (v/v) Triton X-100, and 1 mM dithiothreitol
0.00073
AAUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00046
CAUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00124
GAUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00037
GUUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00081
UAUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00028
UUUUU-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.00032
UUUUU-DN34-containing RNA oligonucleotide
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015
5' hydroxylated fluorogenic oligonucleotide
Escherichia coli
-
pH 7.5, 25°C, recombinant N-terminal domain
-
0.014
5' monophosphorylated fluorogenic oligonucleotide
Escherichia coli
-
pH 7.5, 25°C, recombinant N-terminal domain
-
1.4
5'-GGGACAGUAUUUG-3'
Escherichia coli
-
in 25 mM bis-Tris-propane (pH 8.3), 15 mM MgCl2, 100 mM NaCl, 0.1% (v/v) Triton X-100, and 1 mM dithiothreitol
0.192
AAUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.15
CAUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.765
GAUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.121
GUUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.209
UAUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.015
UUUUU-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
0.023
UUUUU-DN34-containing RNA oligonucleotide
Escherichia coli
-
recombinant enzyme N-terminal half, pH 7.5, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
assay at
7.5 - 7.9
-
assay at
7.6
-
assay at
7.9
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 9.5
-
inactive at pH 6.5 and pH 10.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
30 - 45
-
assay at
45 - 60
-
recombinant enzyme
45 - 75
-
assay at, the cleavage site selection of RNase E/G is temperature-dependent, overview
55
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 85
-
recombinant enzyme
30 - 43
-
assay at, the substrate is more stable at 30°C, overview
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
soluble fraction
Manually annotated by BRENDA team
-
enzyme membrane binding induces changes in secondary protein structure and enzymatic activation by stabilizing the protein-folding state and increasing its binding affinity for ist substrate
-
Manually annotated by BRENDA team
-
RNase E and the other constituents of the RNA degradosome are components of the bacterial cytoskeleton, degradosome components, i.e. RNase E, helicase B, polynucleotide hosphorylase, and enolase, are organized as helical filamentous structures that coil around the length of the cell. Formation of the RNaseE cytoskeletal-like structure requires an internal domain of the protein that does not include the domains required for any of its known interactions or the minimal domain required for endonuclease activity, but is independent of MreB and MinD cytoskeletal structures, mechanism of assembly, overview
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Caulobacter crescentus (strain ATCC 19089 / CB15)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11000
x * 170000, SDS-PAGE, x * 11000, calculated
11936
2 * 11936, calculated from sequence
23700
gel filtration
90000
-
x * 90000, about, SDS-PAGE
115000
-
x * 115000, about, sequence calculation, x * 150000-160000, SDS-PAGE
118000
-
x * 180000, SDS-PAGE, x * 118000, sequence calculation
122600
calculated from sequence
124000 - 132000
-
MW of recombinant dimeric ribonuclease E N-terminal catalytic domains of mutants C404A and C407A by mass spectrometry, gel filtration, and sequence calculations, overview
140000
-
x * 140000, SDS-PAGE
170000
x * 170000, SDS-PAGE, x * 11000, calculated
180000
247500
-
tetrameric wild-type ribonuclease E N-terminal catalytic domain, sequence calculation
248800
-
tetrameric wild-type ribonuclease E N-terminal catalytic domain, mass spectrometry
260000
-
tetrameric catalytic domains, gel filtration
270000
-
about, tetrameric wild-type ribonuclease E N-terminal catalytic domain, gel filtration
300000
-
N-terminal catalytic domain homotetramer, analytical ultracentrifugation
464000
-
co-sedimentation
680000
-
gel filtration, the 680 kDa complex is resistant to a high salt concentration of up to 2 M KCl , but is disrupted by 10 mM EDTA
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oligomer
-
x * 115000, about, sequence calculation, x * 150000-160000, SDS-PAGE
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
-
the recombinant enzyme is cleaved from an active 180 kDa form to active about 70 kDa and 60 kDa fragments during purification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
apoprotein, X-ray diffraction structure determination and analysis at 3.3 resolution, modelling using molecular replacement; RNase E catalytic domain in the apo-state, molecular replacement
-
enolase bound to its cognate site from RNase E, residues 823-850, X-ray diffraction structure determination and analysis at 1.9 A resolution; the crystal structure of enolase bound to its cognate site from RNase E, residues 823-850, at 1.9 A resolution. The structure suggests that enolase may help to organize an adjacent conserved RNA-binding motif in RNase E
-
PNPase complexed with the recognition site from RNase E and with manganese in the presence or in the absence of modified RNA, hanging drop vapour diffusion method, using 0.2 M ammonium nitrate and 20% w/v PEG 3350 or 0.2 M diammonium hydrogen citrate and 17% PEG 3350
-
purified recombinant detagged isolated S1 domain, residues 35-125, large crystals grow within 4 weeks in 1.65 mM protein containing solution of 20 mM phosphate, pH 6.5, 50 mM NaCl, and 0.05% w/v NaN3 at 4°C, isomorphous crystals are grown by hanging drop vapour diffusion method at 18°C, 1.3 mM protein in 20 mM HEPES, p 6.5, 50 mM NaCl, is mixed with a well solution containing 0.17 M sodium acetate, pH 6.5, 85 mM sodium cacodylate, 50% w/v PEG 8000, and 15% glycerol, X-ray diffraction structure determination and analysis at 2.0 A resolution using single anomalous dispersion or trimethyl lead(IV) acetate derivatives
-
purified recombinant His-tagged N-terminal RNaseE catalytic N domain, vapour diffusion method, 7 mg/ml protein in solution is mixed in a 1:1 ratio with precipitation solution containing 0.18 M Li2SO4, 0.09 M Tris-HCl, pH 8.5, 27% w/v PEG 4000, and 10% v/v glycerol, X-ray diffraction structure determination and analysis at 3.4 A resolution
-
X-ray diffraction structure determination and analysis at 2.9 A resolution
-
hanging drop vapor diffusion method. The crystal structure of SSO1404 is solved at 1.6 A resolution revealing the first ribonuclease with a ferredoxin-like fold
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
liposome binding stabilizes protein folding and prevents protein structure changes during thermal denaturation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 20 mM Tris-HCl buffer, pH 7.6, 500 mM NaCl, 10 mM MgCl2, 10 mM DTT, 0.5 mM EDTA, and 5% (v/v) glycerol
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, Toyopearl column filtration chromatography, and Affi-Gel blue column chromatography
-
by affinity chromatography
-
by ammonium sulfate precipitation and cation-exchange chromatography
-
FLAG-tagged RNase E is purified by affinity chromatography
-
immobilized metal affinity chromatography and Superdex 200 gel filtration
-
N-terminal half-RNase E
-
native and recombinant ribonuclease E N-terminal catalytic domains from transformed Escherichia coli
-
native degradosomes from strain CF881, which lacks RNase I, further electrophoretically purification of Rne protein
-
native RNA degradosome from strain BRL2288, recombinant His-tagged full-length wild-type RNase E and His-tagged N-terminal ribonucleolytic domain RTD-RNase E from strain BL21(DE3) by metal affinity chromatography
-
Ni-NTA column chromatography
partially about 100fold by ammonium chloride precipitation, ultracentrifugation, ammonium sulfate fractionation, and gel filtration
-
recombinant active His- and Myc-tagged truncated enzyme from strain BL21(DE3) by affinity chromatography
-
recombinant C-terminally His6- and Myc-tagged N-terminal enzyme half and maltose-binding protein-fused N-terminal half from strain BL21(DE3) by affinity chromatography on a metal chelating resin and an amylose resin, respectively
-
recombinant enzyme from Escherichia coli
-
recombinant enzyme to near homogeneity by SDS-PAGE and renaturation
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
recombinant His-tagged enzyme from strain BL21(DE3)
-
recombinant His-tagged N-terminal catalytic domain from strain BL21(DE3) by nickel affinity chromatography and gel filtration to over 95% purity
-
recombinant His-tagged N-terminal half of RNase E from strain BL21(DE3)
-
recombinant His-tagged N-terminal RNaseE catalytic N domain from strain BL21(DE3) by metal affinity chromatography and gel filtration to homogeneity
-
recombinant His-tagged truncated enzyme mutants from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis and gel filtration
recombinant His-tagged truncated enzyme mutants from strain BL21(DE3) by nickel affinity chromatography, dialysis and gel filtration
-
recombinant His-tagged wild-type and mutant catalytic domains by nickel affinity chromatography
-
recombinant His10-tagged enzyme from Escherichia coli strain BL21(DE3) by His-trap chromatography with or without cleavage of the His-tag by factor Xa; recombinant His10-tagged RNase E/G from Escherichia coli strain BL21(DE3) by metal affinity chromatography, removal of the His-tag by factorXa and dialysis
-
recombinant isolated, His-tagged catalytic domain Rne498 from strain BL21(DE3) by metal affinity chromatography and dialysis to homogeneity
-
recombinant N-terminally His6-tagged wild-type and mutant full-length enzymes, and isolated N-terminally His6-tagged S1 domain by nickel affinity chromatography, removal of the His-tags by thrombin
-
recombinant wild-type and mutant enzymes, purification involves denaturation and renaturation steps due to the poor solubility of the full-length enzyme
-
recombinant wild-type enzyme and truncated mutant enzyme, partially
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparisons, expression in Escherichia coli
-
expressed as a fusion with an N-terminal His6 tag in Escherichia coli strain BL21 (DE3)
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli DG5alpha and BL21(DE3) cells
-
expressed in Escherichia coli DH5alpha cells
-
expressed in Escherichia coli JM109 cells
-
expressed in Escherichia coli strain BZ99
-
expression in wild-type rne cells and in thermosensitive mutant rne-50 and rne-3071 cells, overexpression of truncated 110 kDa mutant enzyme in rne50 cells
-
expression of His-tagged full-length wild-type RNase E and His-tagged N-terminal ribonucleolytic domain RTD-RNase E, residues 1-498, in strain BL21(DE3)
-
expression of His-tagged N-terminal catalytic domain in strain BL21(DE3)
-
expression of His-tagged truncated enzyme mutants in Escherichia coli strain BL21(DE3), complementation of rne null mutation of Escherichia coli strain KSL2000
expression of His-tagged truncated enzyme mutants in strain BL21(DE3), complementation of rne null mutation of strain KSL2000
-
expression of His-tagged wild-type and mutant catalytic domains
-
expression of N-terminal enzyme half, comprising residues 1-499, as C-terminally His6- and Myc-tagged or maltose-binding protein-fused protein in strain BL21(DE3)
-
expression of N-terminally His6-tagged wild-type and mutant full-length enzymes, and of isolated N-terminally His6-tagged S1 domain comprising residues 35-125
-
expression of the His-tagged N-terminal RNaseE catalytic N domain, comprising residues 1-529, in strain BL21(DE3)
-
expression of wild-type and mutant enzyme and N-terminal catalytic domains in Escherichia coli
-
expression of wild-type and mutant enzymes
-
expression of wild-type and mutant enzymes in different modified strains, cell growth curves, overview
-
FLAG-tagged RNase E is expressed in Escherichia coli BL21(DE3) cells
-
fragments of RNase E, either containing the enolase-binding microdomain (residues 833-851), the PNPase-binding site (residues 844-1061) or both (residues 734-1061)
-
gene rne, expression of His-tagged N-terminal half of RNase E in strain BL21(DE3)
-
gene rne, expression of the isolated, His-tagged catalytic domain Rne498, comprising residues 1-498, in strain BL21(DE3)
-
gene rne, expression of wild-type and mutant enzymes in Escherichia coli and in the Saccharomyces cerevisiae two-hybrid system
-
gene rne, overexpression in Escherichia coli, functional complementation of the RNase E-deficient strain CJ1832 of Escherichia coli
-
gene rne, overexpression in strain BL21(DE3)
-
gene rne, overexpression in strain BL21(DE3) as His-tagged enzyme
-
gene rne-1
-
gene rng, overexpression of His10-tagged RNase E/G, Rng, in Escherichia coli strain BL21(DE3); overexpression of His10-tagged enzyme in Escherichia coli strain BL21(DE3)
-
oligohistidine-tagged polypeptides corresponding to the N-terminal half of RNase E with wild-type or mutant sequences
-
overexpressed in Escherichia coli ER2566
-
overexpression of active His- and Myc-tagged truncated enzyme in strain BL21(DE3)
-
overexpression of plasmid-encoded rne gene increases cell doubling time and can lead to plasmid loss or aquisition of mutations that reduce RNase E activity
-
overexpression of wild-type and truncation mutant enzymes in strain Bl21(DE3), expression in the yeast two-hybrid system, overview
-
recombinant expression of FLAG-tagged truncation mutants
-
recombinant overexpression of the His-tagged enzyme in Escherichia coli
-
rne gene amplified and subcloned into pCR-BluntII-TOPO vector to generate pTOPOS14Rne. Expression plasmid pBADS14Rne constructed by excising the rne gene as a 3.3 kb gene Nco I-Pme I fragment from pTOPOS14Rne and inserting the fragment into the medium copy number plasmid pBADlacZ, putting RNase E expression under the control of the arabinose inducible promoter PBAD, and transformed into Escherichia coli AC23 (rne-1). RNase E putative enolase-binding microdomain (residues 885-909) and the putative PNPase-binding microdomain from S14 RNase E (residues 1015-1094) cloned
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A326T
-
random mutagenesis, mutation in the DNase I subdomain, the mutant shows no detectable binding to p23 RNA due to a reduction in the substrate-binding ability
C404A
-
site-directed mutagenesis, mutation of a zinc binding residue, the mutant shows 200fold decreased activity relative to that of the wild-type enzyme for cleaving a 10-mer RNA substrate, and forms a dimer instead of a tetramer
C407A
-
site-directed mutagenesis, mutation of a zinc binding residue, the mutant shows 200fold decreased activity relative to that of the wild-type enzyme for cleaving a 10-mer RNA substrate, and forms a dimer instead of a tetramer
D303N
-
site-directed mutagenesis of a residue located on the surface of the subdomain of RNase E, the mutant shows about 25fold reduced catalytic activity but almost unaltered RNA binding compared to the wild-type enzyme
E R169Q
-
mutant protein is strongly overexpressed with accumulation of proteolytic fragments
F186C
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
G172A
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
G66S
-
site-directed mutagenesis, the mutation leads to a dramatic destabilization of the OB fold of the S1 domain and leads to increased temperature sensitivity of the mutant compared to the wild-type enzyme
I41N
-
random mutagenesis, mutation in the SI subdomain, the mutant shows no detectable binding to p23 RNA due to a reduction in the substrate-binding ability
K106A
-
site-directed mutagenesis, 60% reduced feedback regulation activity compared to the wild-type enzyme
K112A
-
site-directed mutagenesis, 94% reduced feedback regulation activity compared to the wild-type enzyme
K37A
-
site-directed mutagenesis, 94% reduced feedback regulation activity compared to the wild-type enzyme
K38A
-
site-directed mutagenesis, 49% reduced feedback regulation activity compared to the wild-type enzyme
K43A
-
site-directed mutagenesis, 33% reduced feedback regulation activity compared to the wild-type enzyme
K71A
-
site-directed mutagenesis, 56% reduced feedback regulation activity compared to the wild-type enzyme
L112A
-
site-directed mutagenesis of a residue located at the hydrophobic pocket on the surface of the S1 domain, the mutant shows about 50fold reduced catalytic activity compared to the wild-type enzyme
L385P
-
random mutagenesis, mutation in the DNase I subdomain, the mutant shows no detectable binding to p23 RNA due to a reduction in the substrate-binding ability
N305L
-
site-directed mutagenesis of a residue located on the surface of the subdomain of RNase E, the mutant shows reduced catalytic activity compared to the wild-type enzyme
Q36R
-
the mutant is hyperactive in comparison to wild type enzyme. The mutation enhances the RNA binding to the catalytic site of the enzyme
R109A
-
site-directed mutagenesis, 78% reduced feedback regulation activity compared to the wild-type enzyme
R1269Q/DELTA530-1061
-
mutation is lethal
R1269Q/DELTA589-1061
-
mutation is lethal
R1269Q/DELTA730-1061
-
mutant is viable
R169Q
-
site-directed mutagensis, the viable mutation in the 5'-phosphate sensor of RNase E, becomes lethal in combination with deletions removing part of the non-catalytic C-terminal domain of RNase E. Loss of autoregulation in R169Q
R187L
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
R48A
-
site-directed mutagenesis, 49% reduced feedback regulation activity compared to the wild-type enzyme
R64A
-
site-directed mutagenesis, 77% reduced feedback regulation activity compared to the wild-type enzyme
R87A
-
site-directed mutagenesis, 16% increased feedback regulation activity compared to the wild-type enzyme
R95A
-
site-directed mutagenesis, 19% increased feedback regulation activity compared to the wild-type enzyme
T170A
-
site-directed mutagensis, the viable mutation in the 5'-phosphate sensor of RNase E, becomes lethal in combination with deletions removing part of the non-catalytic C-terminal domain of RNase E
T170A/DELTA530-1061
-
mutant is viable, with small colony sizes
T170A/DELTA589-1061
-
mutant is viable, with small colony sizes
Y25A
-
the mutant is hypoactive in comparison to wild type enzyme. The mutation increases the RNA binding to the multimer formation interface between amino acid residues 427 and 433
Y42A
-
site-directed mutagenesis, 48% reduced feedback regulation activity compared to the wild-type enzyme
Y60A
-
site-directed mutagenesis, 99% reduced feedback regulation activity compared to the wild-type enzyme
Y77A
-
site-directed mutagenesis, 19% reduced feedback regulation activity compared to the wild-type enzyme
N305D
-
the mutation is localized in the catalytic domain of RNase E
-
Q36R
-
the mutant is hyperactive in comparison to wild type enzyme. The mutation enhances the RNA binding to the catalytic site of the enzyme
-
Y25A
-
the mutant is hypoactive in comparison to wild type enzyme. The mutation increases the RNA binding to the multimer formation interface between amino acid residues 427 and 433
-
F186C
-
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
-
G172A
-
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
-
R187L
-
site-directed mutagenesis of a point mutation in the S1 RNA-binding domain of RNase E, which leads to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay, and tRNA maturation, intragenic suppressors, rne-172, rne-186 and rne-187 alleles, of the temperature-sensitive rne mutant allele cause the dissociation of RNase E activity on mRNA and tRNA or rRNA substrates in Escherichia coli. Specifically, tRNA maturation and 9S rRNA processing are restored to wild-type levels in suppressor mutants, while mRNA decay remains defective, phenotypes, overview
-
A327P
-
mutation in N-terminal part of enzyme, temperature-sensitive mutation that is able to suppress the slow growth caused by the mutation tufA499 at permissive temperatures. In addition, mutation causes a large increase 503 in rne mRNA steady state levels; site-directed mutagensis, temperature-sensitive mutant
A448V
-
site-directed mutagensis, the mutation causes steric problemes and leads to conformational changes
C471Y
-
site-directed mutagensis, the mutation causes steric problemes and leads to conformational changes
G113D
-
site-directed mutagensis, the mutation causes steric problemes and leads to conformational changes
G66C
-
mutation in N-terminal part of enzyme, temperature-sensitive mutation that is able to suppress the slow growth caused by the mutation tufA499 at permissive temperatures. In addition, mutation causes a large increase 503 in rne mRNA steady state levels; site-directed mutagensis, temperature-sensitive mutant, the mutation causes steric problemes and leads to conformational changes
I207N
-
mutation in N-terminal part of enzyme, temperature-sensitive mutation that is able to suppress the slow growth caused by the mutation tufA499 at permissive temperatures. In addition, mutation causes a large increase 503 in rne mRNA steady state levels; site-directed mutagensis, temperature-sensitive mutant, the mutation reduces the nonpolar contacts in the core, which may lead to a less stable protein
I207S
-
mutation in N-terminal part of enzyme, temperature-sensitive mutation that is able to suppress the slow growth caused by the mutation tufA499 at permissive temperatures. In addition, mutation causes a large increase 503 in rne mRNA steady state levels; site-directed mutagensis, temperature-sensitive mutant, the mutation reduces the nonpolar contacts in the core, which may lead to a less stable protein
L424R
-
site-directed mutagensis, the mutation reduces the nonpolar contacts in the core, which may lead to a less stable protein
V459G
-
site-directed mutagensis, the mutation reduces the nonpolar contacts in the core, which may lead to a less stable protein
D10A
inactive mutant enzyme
D13A
mutant enzyme shows wild-type level activity
D14A
mutant enzyme shows wild-type level activity
D65A
2fold drop in activity compared to wild-type
F37A
inactive mutant enzyme
N18A
mutant enzyme shows wild-type level activity
Q33A
slightly reduced activity
R17A
very low activity
R19A
very low activity
R31A
inactive mutant enzyme
R67A
mutant enzyme shows wild-type level activity
S35A
slightly reduced activity
T12A
2fold drop in activity compared to wild-type
Y34A
slightly reduced activity
Y9A
very low activity
D10A
-
inactive mutant enzyme
-
D13A
-
mutant enzyme shows wild-type level activity
-
R19A
-
very low activity
-
R31A
-
inactive mutant enzyme
-
R67A
-
mutant enzyme shows wild-type level activity
-
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes, purification involves denaturation and renaturation steps due to the poor solubility of the full-length enzyme
-
RNase E is renatured on the Ni-NTA resin by decreasing the concentration of urea to 0.8 M
-
the diluted Rne is dialyzed against two changes of 100 volumes each of 60 mM NH4Cl, 0.1 mM dithiothreitol, and 10% (v/v) glycerol at 4°C for at least 20 h
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
the enzyme can be a model for investigation of the action of site specific nucleases
medicine
Show AA Sequence (4892 entries)
Please use the Sequence Search for a specific query.
LINKS TO OTHER DATABASES (specific for EC-Number 3.1.26.12)