| Autor: |
Khu, Yee-Ling, Koh, Esther, Lim, Siew Pheng, Tan, Yin Hwee, Brenner, Sydney, Lim, Seng Gee, Hong, Wan Jin, Goh, Phuay-Yee |
| Zdroj: |
The Journal of Virology; January 2001, Vol. 75 Issue: 1 p205-214, 10p |
| Abstrakt: |
ABSTRACTInteraction 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. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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