Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and truncated mutant enzymes in Escherichia coli | Neurospora crassa |
Crystallization (Comment) | Organism |
---|---|
purified N-terminally truncated enzyme mutants (FRH-DELTA100 or FRHDELTA114) free or in complex with ADP and/or a single-stranded 13-nucleotide RNA in several conformations, X-ray diffraction structure determination and analysis st 3.1-3.8 A resolution | Neurospora crassa |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of FRH truncated variants beginning at residue 100 or 114 (FRH-DELTA100 or FRHDELTA114) | Neurospora crassa |
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 | Neurospora crassa |
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 | Neurospora crassa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Neurospora crassa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | Neurospora crassa | - |
ADP + phosphate | - |
? | |
ATP + H2O | Neurospora crassa CBS 708.71 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Neurospora crassa 74-OR23-1A | - |
ADP + phosphate | - |
? | |
ATP + H2O | Neurospora crassa DSM 1257 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Neurospora crassa ATCC 24698 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Neurospora crassa FGSC 987 | - |
ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Neurospora crassa | Q1K502 | - |
- |
Neurospora crassa 74-OR23-1A | Q1K502 | - |
- |
Neurospora crassa ATCC 24698 | Q1K502 | - |
- |
Neurospora crassa CBS 708.71 | Q1K502 | - |
- |
Neurospora crassa DSM 1257 | Q1K502 | - |
- |
Neurospora crassa FGSC 987 | Q1K502 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and truncated mutant enzymes from Escherichia coli | Neurospora crassa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | - |
Neurospora crassa | ADP + phosphate | - |
? | |
ATP + H2O | - |
Neurospora crassa CBS 708.71 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Neurospora crassa 74-OR23-1A | ADP + phosphate | - |
? | |
ATP + H2O | - |
Neurospora crassa DSM 1257 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Neurospora crassa ATCC 24698 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Neurospora crassa FGSC 987 | ADP + phosphate | - |
? | |
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa | ? | - |
- |
|
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa CBS 708.71 | ? | - |
- |
|
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa 74-OR23-1A | ? | - |
- |
|
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa DSM 1257 | ? | - |
- |
|
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa ATCC 24698 | ? | - |
- |
|
additional information | interactions of FRH with RNA and ADP, overview | Neurospora crassa FGSC 987 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
frequency-interacting RNA helicase | - |
Neurospora crassa |
FRH | - |
Neurospora crassa |
FRQ-interacting RNA helicase | - |
Neurospora crassa |
General Information | Comment | Organism |
---|---|---|
metabolism | in the Neurospora crassa circadian core clock oscillator, FRH associates with FRQ and CK1a to form a repressor complex (FFC) that inhibits the positive-acting white collar transcription factors (WC-1 and WC-2) which form the white-collar complex (WCC) | Neurospora crassa |
additional information | structure-function analysis of Frh, three-dimensional structure modelling, detailed overview. Structure comparisons to the yeast homologue Mtr4 reveal structure differences and the distinct role of FRH in the clock. The KOW module at an exposed position within a positively charged patch of residues that include K811, K731, R812, K766, R806, K739, R831, R832, and K824. Key residues for FRH function are: K208 and S324 compose the ATP-binding site, E294 lies between the ADP and RNA pockets and participates in ATP hydrolysis, P871 resides in the elbow region of the arch domain. K811, R806, R712, and K766 contribute to a positively charged surface on the KOW domain module. R806H disrupts the clock. Q131, G132, and V142 in the N-terminus provide interactions to the helicase RecA domains. Analysis of interactions analysis of the N-terminal FRQ interaction region, and of interactions of the KOW module with clock components | Neurospora crassa |
physiological function | FRQ-interacting RNA helicase (FRH) is an ATP-dependent RNA helicase that functions in the central transcriptional-translational feedback loop of the Neurospora crassa circadian clock. In the Neurospora crassa circadian clock, a protein complex of frequency (FRQ), casein kinase 1a (CK1a), and the FRQ-interacting RNA helicase (FRH) rhythmically represses gene expression by the white-collar complex (WCC). FRH is essential for cell viability, although FRH helicase activity does not play an essential role in the clock, but rather acts to mediate contacts among FRQ, CK1a and the WCC through interactions involving its N-terminus and KOW module | Neurospora crassa |