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(Received for publication, January 19, 1996, and in revised form, March 21, 1996)
From the We have shown recently that the immunophilins
CyP-40 and FKBP52/hsp56 bind to a common site on hsp90 and that they
exist in separate heterocomplexes with the glucocorticoid receptor
(GR). FKBP52/hsp56 binds to hsp90 via its tetratricopeptide repeat
(TPR) domains, it is not required for GR·hsp90 heterocomplex
assembly, and it is thought to play a role in targeted movement of the
GR. In this work we examine the hsp90 binding of four proteins
(FKBP52/hsp56, CyP-40, p50, Mas70p) thought to be involved in targeted
protein trafficking. FKBP52/hsp56 and CyP-40 (each with three TPRs),
localize to the nucleus and nucleoli, respectively, and form relatively
weak complexes with hsp90 that are competed by a CyP-40 fragment
containing its three TPRs. The p50 component of the Src·hsp90 and
Raf·hsp90 heterocomplexes localizes to cytoskeletal fibers extending
from the perinuclear region to the plasma membrane and forming a rim
under the plasma membrane of endothelial cells. p50, Mas70p (seven
TPRs), which is a receptor for mitochondrial import, and the p60 (six
to eight TPRs) component of the steroid receptor·hsp90 heterocomplex
assembly system bind very tightly to hsp90 in a manner that is not
competed by the CyP-40 fragment. However, bacterially expressed p60
blocks the binding of p50, Mas70p, FKBP52/hsp56, and CyP-40 to purified
hsp90. The data are consistent with binding of all of these proteins to
a site on hsp90 that is a general TPR domain acceptor. Our localization
and binding data are used to develop a model in which proteins that are
chaperoned by hsp90 move as dynamic complexes to their cellular sites
of action, with the TPR-containing protein participating in targeting
the movement of the complexes.
Volume 271, Number 23,
Issue of June 7, 1996
pp. 13468-13475
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
,
,
,
and
Department of Pharmacology, The University
of Michigan Medical School, Ann Arbor, Michigan 48109, the
¶ Department of Pharmacology, Yale University School of Medicine,
New Haven, Connecticut 06510, and the 
Department of
Veterinary Science, Pennsylvania State University, University
Park, Pennsylvania 16802
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