title: Contribution of N- and C-terminal domains to the function of Hsp90 in Saccharomyces cerevisiae.
abstract: The molecular chaperone Hsp90 is a regulatory component of some key signalling proteins in the cytosol of eukaryotic cells. For some of these functions, its interaction with co-chaperones is required. Limited proteolysis defined stable folded units of Hsp90. Both an N-terminal (N210) and a C-terminal (262C) fragment interact with non-native substrate proteins in vitro, but with different specificity and ATP dependence. Here, we analysed the functional properties of these Hsp90 fragments in vivo and in vitro. We determined their influence on the general viability and cell growth of Saccharomyces cerevisiae. Expression of N210 or 262C resulted in a dominant-negative phenotype in several yeast strains tested. Their expression was not toxic, but inhibited cell growth. Further, both were unable to restore viability to Hsp90-depleted cells. In addition, N210 and 262C influence the maturation of Hsp90 substrates, such as the glucocorticoid receptor and pp60v-Src kinase. Specifically, 262C forms partially active chaperone complexes, leading to an arrest of the chaperoned substrate at a certain stage of its maturation cycle. This demonstrates the requirement of a sophisticated and cofactor-regulated interplay between N- and C-terminal activities for Hsp90 function in vivo.
authors.affiliation: Institut für Organische Chemie und Biochemie, Technische Universität München, 85747 Garching, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Scheibel
authors.forename: T
authors.initials: T
authors.lastname: Weikl
authors.forename: R
authors.initials: R
authors.lastname: Rimerman
authors.forename: D
authors.initials: D
authors.lastname: Smith
authors.forename: S
authors.initials: S
authors.lastname: Lindquist
authors.forename: J
authors.initials: J
authors.lastname: Buchner
mesh_terms: D002455:Cell Division / Q000502:physiology; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000502:physiology*; D015688:Oncogene Protein pp60(v-src) / Q000502:physiology; D010446:Peptide Fragments; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011980:Receptors, Progesterone / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000502:physiology*
chemical_list: D018841:HSP90 Heat-Shock Proteins; D010446:Peptide Fragments; D011980:Receptors, Progesterone; D015688:Oncogene Protein pp60(v-src)