title: The beet yellows closterovirus p65 homologue of HSP70 chaperones has ATPase activity associated with its conserved N-terminal domain but does not interact with unfolded protein chains.
abstract: The positive-strand RNA genome of beet yellows closterovirus (BYV) encodes a 65 kDa protein (p65) related to the HSP70 family of cell chaperones. The full-sized BYV p65, and N- and C-terminal fragments, with (His)6 tails, were overexpressed in bacteria and purified by metal-chelate chromatography. Using a polyclonal antiserum raised against the C-terminal fragment of p65, evidence was obtained for expression of the viral protein in planta. Purified recombinant p65 and its N-terminal 40 kDa fragment exhibited Mg2+-dependent ATPase activity in vitro. However, unlike its cellular HSP70 homologues, p65 was unable to bind to denatured protein and its ATPase activity was not stimulated by synthetic peptides which are known to stimulate HSP70 ATPases. Hence, the BYV p65, although being a chaperone-type ATPase, may have a distinct substrate specificity and function in BYV-infected cells.
authors.affiliation: Department of Virology, Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia. AAA@closter.genebee.msu.su
authors.forename: A A
authors.initials: AA
authors.lastname: Agranovsky
authors.forename: S Y
authors.initials: SY
authors.lastname: Folimonova
authors.forename: A S
authors.initials: AS
authors.lastname: Folimonov
authors.forename: O N
authors.initials: ON
authors.lastname: Denisenko
authors.forename: R A
authors.initials: RA
authors.lastname: Zinovkin
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D017790:Closterovirus / Q000378:metabolism*; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C093182:protein p65, Beet yellows virus; D000251:Adenosine Triphosphatases