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. 1998 May;7(5):1186–1194. doi: 10.1002/pro.5560070513

Isolation and characterization of a DnaJ-like protein in rats: the C-terminal 10-kDa domain of hsc70 is not essential for stimulating the ATP-hydrolytic activity of hsc70 by a DnaJ-like protein.

C H Leng 1, J L Brodsky 1, C Wang 1
PMCID: PMC2143995  PMID: 9605323

Abstract

A DnaJ-like protein, RDJ1, was isolated from a rat brain cDNA library. The protein is predicted to have 397 amino acid residues and shares 99% identity to that of HDJ2, a human DnaJ-like protein. RDJ1 was also shown to rescue the temperature-sensitive lethality of a strain containing a mutated cytosolic DnaJ in yeast, ydj1-151. Fragments containing the J-domain of RDJ1 either with or without the G/F motif were expressed in Escherichia coli. The purified proteins stimulated the ATPase activity of hsc70 and of the 60-kDa N-terminal fragment of hsc70. These results imply that RDJ1 can interact with the N-terminal 60-kDa fragment of hsc70 to activate ATP hydrolysis by hsc70.

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Selected References

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  1. Atencio D. P., Yaffe M. P. MAS5, a yeast homolog of DnaJ involved in mitochondrial protein import. Mol Cell Biol. 1992 Jan;12(1):283–291. doi: 10.1128/mcb.12.1.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banecki B., Liberek K., Wall D., Wawrzynów A., Georgopoulos C., Bertoli E., Tanfani F., Zylicz M. Structure-function analysis of the zinc finger region of the DnaJ molecular chaperone. J Biol Chem. 1996 Jun 21;271(25):14840–14848. doi: 10.1074/jbc.271.25.14840. [DOI] [PubMed] [Google Scholar]
  3. Beckmann R. P., Mizzen L. E., Welch W. J. Interaction of Hsp 70 with newly synthesized proteins: implications for protein folding and assembly. Science. 1990 May 18;248(4957):850–854. doi: 10.1126/science.2188360. [DOI] [PubMed] [Google Scholar]
  4. Braun J. E., Wilbanks S. M., Scheller R. H. The cysteine string secretory vesicle protein activates Hsc70 ATPase. J Biol Chem. 1996 Oct 18;271(42):25989–25993. doi: 10.1074/jbc.271.42.25989. [DOI] [PubMed] [Google Scholar]
  5. Caplan A. J., Cyr D. M., Douglas M. G. Eukaryotic homologues of Escherichia coli dnaJ: a diverse protein family that functions with hsp70 stress proteins. Mol Biol Cell. 1993 Jun;4(6):555–563. doi: 10.1091/mbc.4.6.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caplan A. J., Douglas M. G. Characterization of YDJ1: a yeast homologue of the bacterial dnaJ protein. J Cell Biol. 1991 Aug;114(4):609–621. doi: 10.1083/jcb.114.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chappell T. G., Konforti B. B., Schmid S. L., Rothman J. E. The ATPase core of a clathrin uncoating protein. J Biol Chem. 1987 Jan 15;262(2):746–751. [PubMed] [Google Scholar]
  8. Cheetham M. E., Jackson A. P., Anderton B. H. Regulation of 70-kDa heat-shock-protein ATPase activity and substrate binding by human DnaJ-like proteins, HSJ1a and HSJ1b. Eur J Biochem. 1994 Nov 15;226(1):99–107. doi: 10.1111/j.1432-1033.1994.tb20030.x. [DOI] [PubMed] [Google Scholar]
  9. Chellaiah A., Davis A., Mohanakumar T. Cloning of a unique human homologue of the Escherichia coli DNAJ heat shock protein. Biochim Biophys Acta. 1993 Jul 18;1174(1):111–113. doi: 10.1016/0167-4781(93)90103-k. [DOI] [PubMed] [Google Scholar]
  10. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  11. Cyr D. M., Langer T., Douglas M. G. DnaJ-like proteins: molecular chaperones and specific regulators of Hsp70. Trends Biochem Sci. 1994 Apr;19(4):176–181. doi: 10.1016/0968-0004(94)90281-x. [DOI] [PubMed] [Google Scholar]
  12. Flaherty K. M., DeLuca-Flaherty C., McKay D. B. Three-dimensional structure of the ATPase fragment of a 70K heat-shock cognate protein. Nature. 1990 Aug 16;346(6285):623–628. doi: 10.1038/346623a0. [DOI] [PubMed] [Google Scholar]
  13. Flynn G. C., Chappell T. G., Rothman J. E. Peptide binding and release by proteins implicated as catalysts of protein assembly. Science. 1989 Jul 28;245(4916):385–390. doi: 10.1126/science.2756425. [DOI] [PubMed] [Google Scholar]
  14. Freeman B. C., Myers M. P., Schumacher R., Morimoto R. I. Identification of a regulatory motif in Hsp70 that affects ATPase activity, substrate binding and interaction with HDJ-1. EMBO J. 1995 May 15;14(10):2281–2292. doi: 10.1002/j.1460-2075.1995.tb07222.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Frydman J., Nimmesgern E., Ohtsuka K., Hartl F. U. Folding of nascent polypeptide chains in a high molecular mass assembly with molecular chaperones. Nature. 1994 Jul 14;370(6485):111–117. doi: 10.1038/370111a0. [DOI] [PubMed] [Google Scholar]
  16. Georgopoulos C. The emergence of the chaperone machines. Trends Biochem Sci. 1992 Aug;17(8):295–299. doi: 10.1016/0968-0004(92)90439-g. [DOI] [PubMed] [Google Scholar]
  17. Georgopoulos C., Welch W. J. Role of the major heat shock proteins as molecular chaperones. Annu Rev Cell Biol. 1993;9:601–634. doi: 10.1146/annurev.cb.09.110193.003125. [DOI] [PubMed] [Google Scholar]
  18. Hendrick J. P., Langer T., Davis T. A., Hartl F. U., Wiedmann M. Control of folding and membrane translocation by binding of the chaperone DnaJ to nascent polypeptides. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10216–10220. doi: 10.1073/pnas.90.21.10216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hu S. M., Wang C. Involvement of the 10-kDa C-terminal fragment of hsc70 in complexing with unfolded protein. Arch Biochem Biophys. 1996 Aug 1;332(1):163–169. doi: 10.1006/abbi.1996.0328. [DOI] [PubMed] [Google Scholar]
  20. Huang S. P., Tsai M. Y., Tzou Y. M., Wu W. G., Wang C. Aspartyl residue 10 is essential for ATPase activity of rat hsc70. J Biol Chem. 1993 Jan 25;268(3):2063–2068. [PubMed] [Google Scholar]
  21. Langer T., Lu C., Echols H., Flanagan J., Hayer M. K., Hartl F. U. Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding. Nature. 1992 Apr 23;356(6371):683–689. doi: 10.1038/356683a0. [DOI] [PubMed] [Google Scholar]
  22. Liberek K., Marszalek J., Ang D., Georgopoulos C., Zylicz M. Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK. Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2874–2878. doi: 10.1073/pnas.88.7.2874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Minami Y., Höhfeld J., Ohtsuka K., Hartl F. U. Regulation of the heat-shock protein 70 reaction cycle by the mammalian DnaJ homolog, Hsp40. J Biol Chem. 1996 Aug 9;271(32):19617–19624. doi: 10.1074/jbc.271.32.19617. [DOI] [PubMed] [Google Scholar]
  24. Oh S., Iwahori A., Kato S. Human cDNA encoding DnaJ protein homologue. Biochim Biophys Acta. 1993 Jul 18;1174(1):114–116. doi: 10.1016/0167-4781(93)90104-l. [DOI] [PubMed] [Google Scholar]
  25. Ohtsuka K. Cloning of a cDNA for heat-shock protein hsp40, a human homologue of bacterial DnaJ. Biochem Biophys Res Commun. 1993 Nov 30;197(1):235–240. doi: 10.1006/bbrc.1993.2466. [DOI] [PubMed] [Google Scholar]
  26. Palleros D. R., Welch W. J., Fink A. L. Interaction of hsp70 with unfolded proteins: effects of temperature and nucleotides on the kinetics of binding. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5719–5723. doi: 10.1073/pnas.88.13.5719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pellecchia M., Szyperski T., Wall D., Georgopoulos C., Wüthrich K. NMR structure of the J-domain and the Gly/Phe-rich region of the Escherichia coli DnaJ chaperone. J Mol Biol. 1996 Jul 12;260(2):236–250. doi: 10.1006/jmbi.1996.0395. [DOI] [PubMed] [Google Scholar]
  28. Sadis S., Hightower L. E. Unfolded proteins stimulate molecular chaperone Hsc70 ATPase by accelerating ADP/ATP exchange. Biochemistry. 1992 Oct 6;31(39):9406–9412. doi: 10.1021/bi00154a012. [DOI] [PubMed] [Google Scholar]
  29. Schröder H., Langer T., Hartl F. U., Bukau B. DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat-induced protein damage. EMBO J. 1993 Nov;12(11):4137–4144. doi: 10.1002/j.1460-2075.1993.tb06097.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  31. Srinivasan A., McClellan A. J., Vartikar J., Marks I., Cantalupo P., Li Y., Whyte P., Rundell K., Brodsky J. L., Pipas J. M. The amino-terminal transforming region of simian virus 40 large T and small t antigens functions as a J domain. Mol Cell Biol. 1997 Aug;17(8):4761–4773. doi: 10.1128/mcb.17.8.4761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stirling C. J., Rothblatt J., Hosobuchi M., Deshaies R., Schekman R. Protein translocation mutants defective in the insertion of integral membrane proteins into the endoplasmic reticulum. Mol Biol Cell. 1992 Feb;3(2):129–142. doi: 10.1091/mbc.3.2.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Szabo A., Korszun R., Hartl F. U., Flanagan J. A zinc finger-like domain of the molecular chaperone DnaJ is involved in binding to denatured protein substrates. EMBO J. 1996 Jan 15;15(2):408–417. [PMC free article] [PubMed] [Google Scholar]
  34. Szyperski T., Pellecchia M., Wall D., Georgopoulos C., Wüthrich K. NMR structure determination of the Escherichia coli DnaJ molecular chaperone: secondary structure and backbone fold of the N-terminal region (residues 2-108) containing the highly conserved J domain. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11343–11347. doi: 10.1073/pnas.91.24.11343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsai M. Y., Wang C. Uncoupling of peptide-stimulated ATPase and clathrin-uncoating activity in deletion mutant of hsc70. J Biol Chem. 1994 Feb 25;269(8):5958–5962. [PubMed] [Google Scholar]
  36. Wang T. F., Chang J. H., Wang C. Identification of the peptide binding domain of hsc70. 18-Kilodalton fragment located immediately after ATPase domain is sufficient for high affinity binding. J Biol Chem. 1993 Dec 15;268(35):26049–26051. [PubMed] [Google Scholar]
  37. Wawrzynów A., Zylicz M. Divergent effects of ATP on the binding of the DnaK and DnaJ chaperones to each other, or to their various native and denatured protein substrates. J Biol Chem. 1995 Aug 18;270(33):19300–19306. doi: 10.1074/jbc.270.33.19300. [DOI] [PubMed] [Google Scholar]
  38. Yeh H., Lee J., Tsai S., Hsieh C., Tam M. F. Rat kidney glutathione S-transferase 1 subunits have C-terminal truncations. Biochem J. 1996 Mar 15;314(Pt 3):1017–1025. doi: 10.1042/bj3141017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zhang J., Walker G. C. Identification of elements of the peptide binding site of DnaK by peptide cross-linking. J Biol Chem. 1996 Aug 16;271(33):19668–19674. [PubMed] [Google Scholar]
  40. Zhu X., Zhao X., Burkholder W. F., Gragerov A., Ogata C. M., Gottesman M. E., Hendrickson W. A. Structural analysis of substrate binding by the molecular chaperone DnaK. Science. 1996 Jun 14;272(5268):1606–1614. doi: 10.1126/science.272.5268.1606. [DOI] [PMC free article] [PubMed] [Google Scholar]

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