title: Point mutation in calcium-binding domain of mouse polyomavirus VP1 protein does not prevent virus-like particle formation, but changes VP1 interactions with Saccharomyces cerevisiae cell structures.
abstract: The mouse polyomavirus gene for the major structural protein, VP1, with point mutation in the calcium-binding pocket (VP1(Ala)), was expressed in Saccharomyces cerevisiae and in a baculovirus expression system. Surprisingly, VP1(Ala) forms virus-like particles (VLPs) in nuclei of both yeast and insect cells. VP1(Ala)-VLPs produced in S. cerevisiae are unstable and, unlike wild-type VP1 (VP1(wt))-VLPs, they disassemble during the purification procedure and storage. In contrast to VP1(wt), VP1(Ala) does not interact with the yeast mitotic spindle. Nevertheless, both wild-type and mutated VP1 inhibit yeast cell growth. The inhibition is cAMP-dependent. The production of VP1(Ala) and VP1(wt)-VLPs in insect cells also revealed differences in their interactions with cellular protein(s). Thus, the mutation in the VP1 calcium pocket alters the stability and surface conformation of VLPs rather than the ability of VP1 to self-assemble.
authors.affiliation: Department of Genetics and Microbiology, Faculty of Science, Charles University, Vinicná 5, 128 44 Prague 2, Czech Republic.
authors.forename: Tomás
authors.initials: T
authors.lastname: Adamec
authors.forename: Zdena
authors.initials: Z
authors.lastname: Palková
authors.forename: Klára
authors.initials: K
authors.lastname: Velková
authors.forename: Jitka
authors.initials: J
authors.lastname: Stokrová
authors.forename: Jitka
authors.initials: J
authors.lastname: Forstová
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics; D002478:Cells, Cultured; D051379:Mice; D017354:Point Mutation; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism; D018411:Spodoptera; D014404:Tubulin / Q000378:metabolism; D014771:Virion / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D014404:Tubulin; C062481:VP1 protein, polyomavirus; D002118:Calcium