Reconstitution of hepatitis C virus protease activities in yeast

FEBS Letters

Palant, Olga¹  Mak, Paul¹  Plotch, Steve¹  Labonté, Patrick¹ 

[1] Molecular Biology and Virology Section, Wyeth-Ayerst Research, 401 N. Middletown Road, Pearl River, NY 10965, USA

关键词: HCV protease;    Retinoic acid receptor;    Saccharomyces cerevisiae;   

DOI  :  10.1016/S0014-5793(01)02678-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The hepatitis C virus (HCV) protease genes (NS2/3 and NS3) were expressed in yeast with their natural substrates fused to a ligand-dependent transcriptional activator, the retinoic acid receptor (RARβ). RARβ can activate transcription in yeast cells in response to retinoic acids. We hypothesized that cis-cleavage at the NS2–3 or NS3–4A junctions by the appropriate HCV proteases would release RARβ, thereby activating transcription of a reporter gene. Our results from Western blot analyses and reporter gene activation indicate that the wild-type NS2/3 and NS3 enzymes are catalytically active in yeast cells, whereas mutations in the catalytic domain of NS2(C993V) and NS3(S1165A) lead to inactive enzymes. We conclude that HCV NS2/3 and NS3 protease activities can be reconstituted in yeast.

【 授权许可】

Unknown