Interaction between the N-terminal and middle regions is essential for the in vivo function of HSP90 molecular chaperone

Abstract

At the primary structure level, the 90-kDa heat shock protein (HSP90) is composed of three regions: the N-terminal (Met 1-Arg 400), middle (Glu 401-Lys 615), and C-terminal (Asp 621-Asp 732) regions. In the present study, we investigated potential subregion structures of these three regions and their roles. Limited proteolysis revealed that the N-terminal region could be split into two fragments carrying residues Met 1 to Lys 281 (or Lys 283) and Glu 282 (or Tyr 284) to Arg 400. The former is known to carry the ATP-binding domain. The fragments carrying the N-terminal two-thirds (Glu 401-Lys 546) and C-terminal one-third of the middle region were sufficient for the interactions with the N- and C-terminal regions, respectively. Yeast HSC82 that carried point mutations in the middle region causing deficient binding to the N-terminal region could not support the growth of HSP82-depleted cells at an elevated temperature. Taken together, our data show that the N-terminal and middle regions of the HSP90 family protein are structurally divided into two respective subregions. Moreover, the interaction between the N-terminal and middle regions is essential for the in vivo function of HSP90 in yeast.