title: Survey of molecular chaperone requirement for the biosynthesis of hamster polyomavirus VP1 protein in Saccharomyces cerevisiae.
abstract: A number of viruses utilize molecular chaperones during various stages of their life cycle. It has been shown that members of the heat-shock protein 70 (Hsp70) chaperone family assist polyomavirus capsids during infection. However, the molecular chaperones that assist the formation of recombinant capsid viral protein 1 (VP1)-derived virus-like particles (VLPs) in yeast remain unclear. A panel of yeast strains with single chaperone gene deletions were used to evaluate the chaperones required for biosynthesis of recombinant hamster polyomavirus capsid protein VP1. The impact of deletion or mild overexpression of chaperone genes was determined in live cells by flow cytometry using enhanced green fluorescent protein (EGFP) fused with VP1. Targeted genetic analysis demonstrated that VP1-EGFP fusion protein levels were significantly higher in yeast strains in which the SSZ1 or ZUO1 genes encoding ribosome-associated complex components were deleted. The results confirmed the participation of cytosolic Hsp70 chaperones and suggested the potential involvement of the Ydj1 and Caj1 co-chaperones and the endoplasmic reticulum chaperones in the biosynthesis of VP1 VLPs in yeast. Likewise, the markedly reduced levels of VP1-EGFP in Δhsc82 and Δhsp82 yeast strains indicated that both Hsp70 and Hsp90 chaperones might assist VP1 VLPs during protein biosynthesis.
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241, Vilnius, Lithuania.
authors.forename: Monika
authors.initials: M
authors.lastname: Valaviciute
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241, Vilnius, Lithuania.
authors.forename: Milda
authors.initials: M
authors.lastname: Norkiene
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241, Vilnius, Lithuania.
authors.forename: Karolis
authors.initials: K
authors.lastname: Goda
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241, Vilnius, Lithuania.
authors.forename: Rimantas
authors.initials: R
authors.lastname: Slibinskas
authors.affiliation: Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241, Vilnius, Lithuania. agedv@ibt.lt.
authors.forename: Alma
authors.initials: A
authors.lastname: Gedvilaite
mesh_terms: D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae / Q000821:virology; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D011120:Polyomavirus / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C088621:CAJ1 protein, S cerevisiae; D002148:Calmodulin-Binding Proteins; D036022:Capsid Proteins; D050956:HSP40 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; C070319:YDJ1 protein, S cerevisiae