PMID:9233823

Citation	Carles-Kinch, K, George, JW and Kreuzer, KN (1997) Bacteriophage T4 UvsW protein is a helicase involved in recombination, repair and the regulation of DNA replication origins. EMBO J. 16:4142-51
Abstract	Bacteriophage T4 UvsW protein is involved in phage recombination, repair and the regulation of replication origins. Here, we provide evidence that UvsW functions as a helicase. First, expression of UvsW allows growth of an (otherwise inviable) Escherichia coli recG rnhA double mutant, consistent with UvsW being a functional analog of the RecG helicase. Second, UvsW contains helicase sequence motifs, and a substitution (K141R) in the Walker 'A' motif prevents growth of the E.coli recG rnhA double mutant. Third, UvsW, but not UvsW-K141R, inhibits replication from a T4 origin at which persistent RNA-DNA hybrids form and presumably trigger replication initiation. Fourth, mutations that inactivate UvsW and endonuclease VII (which cleaves DNA branches) synergistically block repair of double-strand breaks. These in vivo results are consistent with a model in which UvsW is a DNA helicase that catalyzes branch migration and dissociation of RNA-DNA hybrids. In support of this model, a partially purified GST/UvsW fusion protein, but not a GST/UvsW-K141R fusion, displays ssDNA-dependent ATPase activity and is able to unwind a branched DNA substrate.
Links	PubMed PMC1170037 Online version:10.1093/emboj/16.13.4142
Keywords	Adenosine Triphosphatases/metabolism; Bacterial Proteins/genetics; Bacteriophage T4/enzymology; Bacteriophage T4/genetics; Bacteriophage T4/physiology; DNA/metabolism; DNA Helicases/genetics; DNA Helicases/metabolism; DNA Repair; DNA, Single-Stranded; Endodeoxyribonucleases/genetics; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/virology; Escherichia coli Proteins; Glutathione Transferase/genetics; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Replication Origin; Ribonuclease H/genetics; Viral Proteins/genetics; Viral Proteins/metabolism; Virus Replication
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9CAUD:S5YPI0	enables	GO:0004386: helicase activity	ECO:0000315: mutant phenotype evidence used in manual assertion		F		Seeded From UniProt	complete
9CAUD:S5YPI0		GO:0004386: helicase activity	ECO:0000315:		F		UvsW acts as an Helicase in T4-bacteriophage, where it functions to unwind DNA/RNA double strands and suppress the R loops in order for the replication process to take place. Table 1. results show that when a UvsW protein was inserted into a E.coli double mutant, which lacked rnhA protein and caused formation of R loops and lead to cell death, it suppressed the R loop formation.	complete CACAO 10885
BPT4:UVSW		GO:0008026: ATP-dependent helicase activity	ECO:0000314:		F		In figure 6, y-branched linear dsDNA is incubated with a GST/uvsW fusion protein and single-stranded products are observed, indicating the unwinding (i.e., helicase) activity of the protein. In lane 9 of the gel, removal of ATP from the reaction mixture abolishes the observed unwinding activity.	complete CACAO 11992

References

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