Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Sep 15;90(18):8557–8561. doi: 10.1073/pnas.90.18.8557

Isoprenylation of the plant molecular chaperone ANJ1 facilitates membrane association and function at high temperature.

J K Zhu 1, R A Bressan 1, P M Hasegawa 1
PMCID: PMC47396  PMID: 8378331

Abstract

We demonstrate that ANJ1, a higher plant homolog of the bacterial molecular chaperone DnaJ, is a substrate in vitro for protein farnesyl- and geranylgeranyl-transferase activities present in cell extracts of the plant Atriplex nummularia and yeast Saccharomyces cerevisiae. Isoprenylation did not occur when cysteine was replaced by serine in the CAQQ motif at the carboxyl terminus of ANJ1, indicating that this sequence functions as a CaaX consensus sequence for polyisoprenylation (where C is cysteine, a is an aliphatic residue, and X is any amino acid residue). Substitution of leucine for the terminal glutamine did not result in the expected geranylgeranylation as occurs with mammalian proteins containing a carboxyl-terminal leucine. Unlike the wild-type ANJ1, neither of the proteins containing these amino acid substitutions could functionally complement the yeast temperature-sensitive mutant mas5. Farnesylation enhanced the association of ANJ1 with A. nummularia microsomal membranes. Electrophoretic mobility of ANJ1 from the plant indicated that the protein is isoprenylated in vivo.

Full text

PDF
8557

Images in this article

8558Fig. 1
on p.8558
8559Fig. 2
on p.8559
8559Fig. 3
on p.8559
8560Fig. 5
on p.8560
8560Fig. 6
on p.8560
8560Fig. 7
on p.8560

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anant J. S., Ong O. C., Xie H. Y., Clarke S., O'Brien P. J., Fung B. K. In vivo differential prenylation of retinal cyclic GMP phosphodiesterase catalytic subunits. J Biol Chem. 1992 Jan 15;267(2):687–690. [PubMed] [Google Scholar]
  2. Anderegg R. J., Betz R., Carr S. A., Crabb J. W., Duntze W. Structure of Saccharomyces cerevisiae mating hormone a-factor. Identification of S-farnesyl cysteine as a structural component. J Biol Chem. 1988 Dec 5;263(34):18236–18240. [PubMed] [Google Scholar]
  3. 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]
  4. Black S. D. Development of hydrophobicity parameters for prenylated proteins. Biochem Biophys Res Commun. 1992 Aug 14;186(3):1437–1442. doi: 10.1016/s0006-291x(05)81567-0. [DOI] [PubMed] [Google Scholar]
  5. Caplan A. J., Cyr D. M., Douglas M. G. YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism. Cell. 1992 Dec 24;71(7):1143–1155. doi: 10.1016/s0092-8674(05)80063-7. [DOI] [PubMed] [Google Scholar]
  6. Caplan A. J., Tsai J., Casey P. J., Douglas M. G. Farnesylation of YDJ1p is required for function at elevated growth temperatures in Saccharomyces cerevisiae. J Biol Chem. 1992 Sep 15;267(26):18890–18895. [PubMed] [Google Scholar]
  7. Clarke S. Protein isoprenylation and methylation at carboxyl-terminal cysteine residues. Annu Rev Biochem. 1992;61:355–386. doi: 10.1146/annurev.bi.61.070192.002035. [DOI] [PubMed] [Google Scholar]
  8. Farnsworth C. C., Kawata M., Yoshida Y., Takai Y., Gelb M. H., Glomset J. A. C terminus of the small GTP-binding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6196–6200. doi: 10.1073/pnas.88.14.6196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fukada Y., Takao T., Ohguro H., Yoshizawa T., Akino T., Shimonishi Y. Farnesylated gamma-subunit of photoreceptor G protein indispensable for GTP-binding. Nature. 1990 Aug 16;346(6285):658–660. doi: 10.1038/346658a0. [DOI] [PubMed] [Google Scholar]
  10. Glenn J. S., Watson J. A., Havel C. M., White J. M. Identification of a prenylation site in delta virus large antigen. Science. 1992 May 29;256(5061):1331–1333. doi: 10.1126/science.1598578. [DOI] [PubMed] [Google Scholar]
  11. Goldstein J. L., Brown M. S. Regulation of the mevalonate pathway. Nature. 1990 Feb 1;343(6257):425–430. doi: 10.1038/343425a0. [DOI] [PubMed] [Google Scholar]
  12. Hancock J. F., Paterson H., Marshall C. J. A polybasic domain or palmitoylation is required in addition to the CAAX motif to localize p21ras to the plasma membrane. Cell. 1990 Oct 5;63(1):133–139. doi: 10.1016/0092-8674(90)90294-o. [DOI] [PubMed] [Google Scholar]
  13. Holtz D., Tanaka R. A., Hartwig J., McKeon F. The CaaX motif of lamin A functions in conjunction with the nuclear localization signal to target assembly to the nuclear envelope. Cell. 1989 Dec 22;59(6):969–977. doi: 10.1016/0092-8674(89)90753-8. [DOI] [PubMed] [Google Scholar]
  14. Inglese J., Koch W. J., Caron M. G., Lefkowitz R. J. Isoprenylation in regulation of signal transduction by G-protein-coupled receptor kinases. Nature. 1992 Sep 10;359(6391):147–150. doi: 10.1038/359147a0. [DOI] [PubMed] [Google Scholar]
  15. Khosravi-Far R., Lutz R. J., Cox A. D., Conroy L., Bourne J. R., Sinensky M., Balch W. E., Buss J. E., Der C. J. Isoprenoid modification of rab proteins terminating in CC or CXC motifs. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6264–6268. doi: 10.1073/pnas.88.14.6264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kinsella B. T., Erdman R. A., Maltese W. A. Posttranslational modification of Ha-ras p21 by farnesyl versus geranylgeranyl isoprenoids is determined by the COOH-terminal amino acid. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8934–8938. doi: 10.1073/pnas.88.20.8934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kinsella B. T., Maltese W. A. rab GTP-binding proteins with three different carboxyl-terminal cysteine motifs are modified in vivo by 20-carbon isoprenoids. J Biol Chem. 1992 Feb 25;267(6):3940–3945. [PubMed] [Google Scholar]
  18. Kitten G. T., Nigg E. A. The CaaX motif is required for isoprenylation, carboxyl methylation, and nuclear membrane association of lamin B2. J Cell Biol. 1991 Apr;113(1):13–23. doi: 10.1083/jcb.113.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Newman C. M., Giannakouros T., Hancock J. F., Fawell E. H., Armstrong J., Magee A. I. Post-translational processing of Schizosaccharomyces pombe YPT proteins. J Biol Chem. 1992 Jun 5;267(16):11329–11336. [PubMed] [Google Scholar]
  21. Ohki R., Kawamata T., Katoh Y., Hosoda F., Ohki M. Escherichia coli dnaJ deletion mutation results in loss of stability of a positive regulator, CRP. J Biol Chem. 1992 Jul 5;267(19):13180–13184. [PubMed] [Google Scholar]
  22. Philips M. R., Pillinger M. H., Staud R., Volker C., Rosenfeld M. G., Weissmann G., Stock J. B. Carboxyl methylation of Ras-related proteins during signal transduction in neutrophils. Science. 1993 Feb 12;259(5097):977–980. doi: 10.1126/science.8438158. [DOI] [PubMed] [Google Scholar]
  23. Polson A., von Wechmar M. B., van Regenmortel M. H. Isolation of viral IgY antibodies from yolks of immunized hens. Immunol Commun. 1980;9(5):475–493. doi: 10.3109/08820138009066010. [DOI] [PubMed] [Google Scholar]
  24. Randall S. K., Marshall M. S., Crowell D. N. Protein isoprenylation in suspension-cultured tobacco cells. Plant Cell. 1993 Apr;5(4):433–442. doi: 10.1105/tpc.5.4.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rine J., Kim S. H. A role for isoprenoid lipids in the localization and function of an oncoprotein. New Biol. 1990 Mar;2(3):219–226. [PubMed] [Google Scholar]
  26. Song C. S., Yu J. H., Bai D. H., Hester P. Y., Kim K. H. Antibodies to the alpha-subunit of insulin receptor from eggs of immunized hens. J Immunol. 1985 Nov;135(5):3354–3359. [PubMed] [Google Scholar]
  27. Stimmel J. B., Deschenes R. J., Volker C., Stock J., Clarke S. Evidence for an S-farnesylcysteine methyl ester at the carboxyl terminus of the Saccharomyces cerevisiae RAS2 protein. Biochemistry. 1990 Oct 16;29(41):9651–9659. doi: 10.1021/bi00493a021. [DOI] [PubMed] [Google Scholar]
  28. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  29. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  30. Sunshine M., Feiss M., Stuart J., Yochem J. A new host gene (groPC) necessary for lambda DNA replication. Mol Gen Genet. 1977 Feb 28;151(1):27–34. doi: 10.1007/BF00446909. [DOI] [PubMed] [Google Scholar]
  31. Swiezewska E., Thelin A., Dallner G., Andersson B., Ernster L. Occurrence of prenylated proteins in plant cells. Biochem Biophys Res Commun. 1993 Apr 15;192(1):161–166. doi: 10.1006/bbrc.1993.1395. [DOI] [PubMed] [Google Scholar]
  32. Vorburger K., Kitten G. T., Nigg E. A. Modification of nuclear lamin proteins by a mevalonic acid derivative occurs in reticulocyte lysates and requires the cysteine residue of the C-terminal CXXM motif. EMBO J. 1989 Dec 20;8(13):4007–4013. doi: 10.1002/j.1460-2075.1989.tb08583.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yang Z., Cramer C. L., Watson J. C. Protein farnesyltransferase in plants. Molecular cloning and expression of a homolog of the beta subunit from the garden pea. Plant Physiol. 1993 Feb;101(2):667–674. doi: 10.1104/pp.101.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zhu J. K., Shi J., Bressan R. A., Hasegawa P. M. Expression of an Atriplex nummularia gene encoding a protein homologous to the bacterial molecular chaperone DnaJ. Plant Cell. 1993 Mar;5(3):341–349. doi: 10.1105/tpc.5.3.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zylicz M., Ang D., Liberek K., Georgopoulos C. Initiation of lambda DNA replication with purified host- and bacteriophage-encoded proteins: the role of the dnaK, dnaJ and grpE heat shock proteins. EMBO J. 1989 May;8(5):1601–1608. doi: 10.1002/j.1460-2075.1989.tb03544.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES