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. 2000 Feb 1;345(Pt 3):645–651. doi: 10.1042/0264-6021:3450645

Heat shock cognate protein 70 chaperone-binding site in the co-chaperone murine stress-inducible protein 1 maps to within three consecutive tetratricopeptide repeat motifs.

J Van Der Spuy 1, B D Kana 1, H W Dirr 1, G L Blatch 1
PMCID: PMC1220800  PMID: 10642524

Abstract

Murine stress-inducible protein 1 (mSTI1) is a co-chaperone homologous with the human heat shock cognate protein 70 (hsc70)/heat shock protein 90 (hsp90)-organizing protein (Hop). The concomitant interaction of mSTI1 with hsp70 and hsp90 at its N- and C-termini respectively is mediated by the tetratricopeptide repeat (TPR) motifs in these regions. With the use of co-precipitation assays, we show here that the N-terminal TPR domain of mSTI1 without extensive flanking regions is both necessary and sufficient to mediate a specific interaction with hsc70. In contrast, other TPR-containing co-chaperones require TPR flanking regions for target substrate recognition, suggesting different mechanisms of TPR-mediated chaperone-co-chaperone interactions. Furthermore, the interaction between mSTI1 and hsc70 was analysed to ascertain the effect of replacing or deleting conserved amino acid residues and sequences within the three TPR motifs constituting the N-terminal TPR domain of full-length mSTI1. Replacement of a bulky hydrophobic residue in TPR1 disrupted the interaction of mSTI1 with hsc70. A highly conserved sequence in TPR2 was altered by deletion or single amino acid replacement. These derivatives retained a specific interaction with hsc70. These results are consistent with a model in which conserved residues within the N-terminal TPR region of mSTI1 contribute differentially to the interaction with hsc70, and in which TPR1 has a significant role in targeting mSTI1 to hsc70. The contribution of the TPR domain mutations and deletions are discussed with respect to their effect on target substrate interactions.

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Selected References

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  1. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. doi: 10.1093/nar/25.17.3389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barent R. L., Nair S. C., Carr D. C., Ruan Y., Rimerman R. A., Fulton J., Zhang Y., Smith D. F. Analysis of FKBP51/FKBP52 chimeras and mutants for Hsp90 binding and association with progesterone receptor complexes. Mol Endocrinol. 1998 Mar;12(3):342–354. doi: 10.1210/mend.12.3.0075. [DOI] [PubMed] [Google Scholar]
  3. Blatch G. L., Lässle M., Zetter B. R., Kundra V. Isolation of a mouse cDNA encoding mSTI1, a stress-inducible protein containing the TPR motif. Gene. 1997 Jul 31;194(2):277–282. doi: 10.1016/s0378-1119(97)00206-0. [DOI] [PubMed] [Google Scholar]
  4. Carrello A., Ingley E., Minchin R. F., Tsai S., Ratajczak T. The common tetratricopeptide repeat acceptor site for steroid receptor-associated immunophilins and hop is located in the dimerization domain of Hsp90. J Biol Chem. 1999 Jan 29;274(5):2682–2689. doi: 10.1074/jbc.274.5.2682. [DOI] [PubMed] [Google Scholar]
  5. Chang H. C., Nathan D. F., Lindquist S. In vivo analysis of the Hsp90 cochaperone Sti1 (p60). Mol Cell Biol. 1997 Jan;17(1):318–325. doi: 10.1128/mcb.17.1.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen M. S., Silverstein A. M., Pratt W. B., Chinkers M. The tetratricopeptide repeat domain of protein phosphatase 5 mediates binding to glucocorticoid receptor heterocomplexes and acts as a dominant negative mutant. J Biol Chem. 1996 Dec 13;271(50):32315–32320. doi: 10.1074/jbc.271.50.32315. [DOI] [PubMed] [Google Scholar]
  7. Chen S., Prapapanich V., Rimerman R. A., Honoré B., Smith D. F. Interactions of p60, a mediator of progesterone receptor assembly, with heat shock proteins hsp90 and hsp70. Mol Endocrinol. 1996 Jun;10(6):682–693. doi: 10.1210/mend.10.6.8776728. [DOI] [PubMed] [Google Scholar]
  8. Chen S., Smith D. F. Hop as an adaptor in the heat shock protein 70 (Hsp70) and hsp90 chaperone machinery. J Biol Chem. 1998 Dec 25;273(52):35194–35200. doi: 10.1074/jbc.273.52.35194. [DOI] [PubMed] [Google Scholar]
  9. Das A. K., Cohen P. W., Barford D. The structure of the tetratricopeptide repeats of protein phosphatase 5: implications for TPR-mediated protein-protein interactions. EMBO J. 1998 Mar 2;17(5):1192–1199. doi: 10.1093/emboj/17.5.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Demand J., Lüders J., Höhfeld J. The carboxy-terminal domain of Hsc70 provides binding sites for a distinct set of chaperone cofactors. Mol Cell Biol. 1998 Apr;18(4):2023–2028. doi: 10.1128/mcb.18.4.2023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dittmar K. D., Demady D. R., Stancato L. F., Krishna P., Pratt W. B. Folding of the glucocorticoid receptor by the heat shock protein (hsp) 90-based chaperone machinery. The role of p23 is to stabilize receptor.hsp90 heterocomplexes formed by hsp90.p60.hsp70. J Biol Chem. 1997 Aug 22;272(34):21213–21220. doi: 10.1074/jbc.272.34.21213. [DOI] [PubMed] [Google Scholar]
  12. Dittmar K. D., Hutchison K. A., Owens-Grillo J. K., Pratt W. B. Reconstitution of the steroid receptor.hsp90 heterocomplex assembly system of rabbit reticulocyte lysate. J Biol Chem. 1996 May 31;271(22):12833–12839. doi: 10.1074/jbc.271.22.12833. [DOI] [PubMed] [Google Scholar]
  13. Dittmar K. D., Pratt W. B. Folding of the glucocorticoid receptor by the reconstituted Hsp90-based chaperone machinery. The initial hsp90.p60.hsp70-dependent step is sufficient for creating the steroid binding conformation. J Biol Chem. 1997 May 16;272(20):13047–13054. doi: 10.1074/jbc.272.20.13047. [DOI] [PubMed] [Google Scholar]
  14. Gebauer M., Zeiner M., Gehring U. Proteins interacting with the molecular chaperone hsp70/hsc70: physical associations and effects on refolding activity. FEBS Lett. 1997 Nov 3;417(1):109–113. doi: 10.1016/s0014-5793(97)01267-2. [DOI] [PubMed] [Google Scholar]
  15. Goebl M., Yanagida M. The TPR snap helix: a novel protein repeat motif from mitosis to transcription. Trends Biochem Sci. 1991 May;16(5):173–177. doi: 10.1016/0968-0004(91)90070-c. [DOI] [PubMed] [Google Scholar]
  16. Hoffmann K., Handschumacher R. E. Cyclophilin-40: evidence for a dimeric complex with hsp90. Biochem J. 1995 Apr 1;307(Pt 1):5–8. doi: 10.1042/bj3070005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Irmer H., Höhfeld J. Characterization of functional domains of the eukaryotic co-chaperone Hip. J Biol Chem. 1997 Jan 24;272(4):2230–2235. doi: 10.1074/jbc.272.4.2230. [DOI] [PubMed] [Google Scholar]
  18. Johnson B. D., Schumacher R. J., Ross E. D., Toft D. O. Hop modulates Hsp70/Hsp90 interactions in protein folding. J Biol Chem. 1998 Feb 6;273(6):3679–3686. doi: 10.1074/jbc.273.6.3679. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Liu F. H., Wu S. J., Hu S. M., Hsiao C. D., Wang C. Specific interaction of the 70-kDa heat shock cognate protein with the tetratricopeptide repeats. J Biol Chem. 1999 Nov 26;274(48):34425–34432. doi: 10.1074/jbc.274.48.34425. [DOI] [PubMed] [Google Scholar]
  21. Lässle M., Blatch G. L., Kundra V., Takatori T., Zetter B. R. Stress-inducible, murine protein mSTI1. Characterization of binding domains for heat shock proteins and in vitro phosphorylation by different kinases. J Biol Chem. 1997 Jan 17;272(3):1876–1884. doi: 10.1074/jbc.272.3.1876. [DOI] [PubMed] [Google Scholar]
  22. Owens-Grillo J. K., Czar M. J., Hutchison K. A., Hoffmann K., Perdew G. H., Pratt W. B. A model of protein targeting mediated by immunophilins and other proteins that bind to hsp90 via tetratricopeptide repeat domains. J Biol Chem. 1996 Jun 7;271(23):13468–13475. doi: 10.1074/jbc.271.23.13468. [DOI] [PubMed] [Google Scholar]
  23. Peitsch M. C. ProMod and Swiss-Model: Internet-based tools for automated comparative protein modelling. Biochem Soc Trans. 1996 Feb;24(1):274–279. doi: 10.1042/bst0240274. [DOI] [PubMed] [Google Scholar]
  24. Prapapanich V., Chen S., Toran E. J., Rimerman R. A., Smith D. F. Mutational analysis of the hsp70-interacting protein Hip. Mol Cell Biol. 1996 Nov;16(11):6200–6207. doi: 10.1128/mcb.16.11.6200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Prodromou C., Siligardi G., O'Brien R., Woolfson D. N., Regan L., Panaretou B., Ladbury J. E., Piper P. W., Pearl L. H. Regulation of Hsp90 ATPase activity by tetratricopeptide repeat (TPR)-domain co-chaperones. EMBO J. 1999 Feb 1;18(3):754–762. doi: 10.1093/emboj/18.3.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Radanyi C., Chambraud B., Baulieu E. E. The ability of the immunophilin FKBP59-HBI to interact with the 90-kDa heat shock protein is encoded by its tetratricopeptide repeat domain. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11197–11201. doi: 10.1073/pnas.91.23.11197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ratajczak T., Carrello A. Cyclophilin 40 (CyP-40), mapping of its hsp90 binding domain and evidence that FKBP52 competes with CyP-40 for hsp90 binding. J Biol Chem. 1996 Feb 9;271(6):2961–2965. doi: 10.1074/jbc.271.6.2961. [DOI] [PubMed] [Google Scholar]
  28. Russell L. C., Whitt S. R., Chen M. S., Chinkers M. Identification of conserved residues required for the binding of a tetratricopeptide repeat domain to heat shock protein 90. J Biol Chem. 1999 Jul 16;274(29):20060–20063. doi: 10.1074/jbc.274.29.20060. [DOI] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sikorski R. S., Boguski M. S., Goebl M., Hieter P. A repeating amino acid motif in CDC23 defines a family of proteins and a new relationship among genes required for mitosis and RNA synthesis. Cell. 1990 Jan 26;60(2):307–317. doi: 10.1016/0092-8674(90)90745-z. [DOI] [PubMed] [Google Scholar]
  31. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  32. Smith D. F. Sequence motifs shared between chaperone components participating in the assembly of progesterone receptor complexes. Biol Chem. 1998 Mar;379(3):283–288. doi: 10.1515/bchm.1998.379.3.283. [DOI] [PubMed] [Google Scholar]
  33. Smith D. F., Sullivan W. P., Marion T. N., Zaitsu K., Madden B., McCormick D. J., Toft D. O. Identification of a 60-kilodalton stress-related protein, p60, which interacts with hsp90 and hsp70. Mol Cell Biol. 1993 Feb;13(2):869–876. doi: 10.1128/mcb.13.2.869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Young J. C., Obermann W. M., Hartl F. U. Specific binding of tetratricopeptide repeat proteins to the C-terminal 12-kDa domain of hsp90. J Biol Chem. 1998 Jul 17;273(29):18007–18010. doi: 10.1074/jbc.273.29.18007. [DOI] [PubMed] [Google Scholar]

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