The middle domain of Hsp90 acts as a discriminator between different types of client proteins.
The mechanism of client protein activation by Hsp90 is enigmatic, and it is uncertain whether Hsp90 employs a common route for all proteins. Using a mutational analysis approach, we investigated the activation of two types of client proteins, glucocorticoid receptor (GR) and the kinase v-Src by the middle domain of Hsp90 (Hsp90M) in vivo. Remarkably, the overall cellular activity of v-Src was highly elevated in a W300A mutant yeast strain due to a 10-fold increase in cellular protein levels of the kinase. In contrast, the cellular activity of GR remained almost unaffected by the W300A mutation but was dramatically sensitive to S485Y and T525I exchanges. In addition, we show that mutations S485Y and T525I in Hsp90M reduce the ATP hydrolysis rate, suggesting that Hsp90 ATPase is more tightly regulated than assumed previously. Therefore, the activation of GR and v-Src has various demands on Hsp90 biochemistry and is dependent on separate functional regions of Hsp90M. Thus, Hsp90M seems to discriminate between different substrate types and to adjust the molecular chaperone for proper substrate activation.
Hawle
Patricija
P
Protein Folding Group, Institute for Genetics, University of Bonn, Römerstr. 164, Bonn, Germany.
Siepmann
Martin
M
Harst
Anja
A
Siderius
Marco
M
Reusch
H Peter
HP
Obermann
Wolfgang M J
WM
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D002478:Cells, Cultured; D018833:Chaperonins; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism*; D006868:Hydrolysis; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism*; D015688:Oncogene Protein pp60(v-src); D017354:Point Mutation; D017434:Protein Structure, Tertiary; D011965:Receptors, Glucocorticoid / Q000378:metabolism / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology; D015003:Yeasts / Q000166:cytology
C475092:AHA1 protein, S cerevisiae; D005656:Fungal Proteins; D018841:HSP90 Heat-Shock Proteins; D018832:Molecular Chaperones; D011965:Receptors, Glucocorticoid; D029701:Saccharomyces cerevisiae Proteins; D000255:Adenosine Triphosphate; D015688:Oncogene Protein pp60(v-src); D000251:Adenosine Triphosphatases; D018833:Chaperonins