PMID:11119590

Citation	Khu, YL, Koh, E, Lim, SP, Tan, YH, Brenner, S, Lim, SG, Hong, WJ and Goh, PY (2001) Mutations that affect dimer formation and helicase activity of the hepatitis C virus helicase. J. Virol. 75:205-14
Abstract	Interaction between viral proteins is necessary for viral replication and viral particle assembly. We used the yeast two-hybrid assay to identify interactions among all the mature proteins of the hepatitis C virus. The interaction between NS3 and NS3 was one of the strongest viral protein-protein interactions detected. The minimal region required for this interaction was mapped to a specific subdomain of 174 amino acids in the N terminus of the helicase region. Random mutations in the minimal region were generated by PCR, and mutants that failed to interact with a wild-type minimal fragment were isolated using the yeast two-hybrid assay as a screen. Three of these mutations resulted in a reduction or a loss of interaction between helicases. Analytical gel filtration showed that in the presence of an oligonucleotide, wild-type helicases form dimers whereas the mutants remain mostly monomeric. All three mutants were partially or almost inactive when assayed for helicase activity in vitro. Mixing a mutant helicase (Y267S) with wild-type helicase did not dramatically affect helicase activity. These data indicate that dimerization of the helicase is important for helicase activity. The mutations that reduce self-association of the helicase may define the key residues involved in NS3-NS3 dimerization.
Links	PubMed PMC113914 Online version:10.1128/JVI.75.1.205-214.2001
Keywords	Amino Acid Sequence; Animals; COS Cells; Dimerization; Molecular Sequence Data; Mutation; RNA Helicases/chemistry; RNA Helicases/metabolism; Viral Nonstructural Proteins/chemistry; Viral Nonstructural Proteins/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HCVH:POLG	enables	GO:0042802: identical protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P27958:PRO_0000037573	F		Seeded From UniProt	complete

References

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