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. 1993 Feb;101(2):667–674. doi: 10.1104/pp.101.2.667

Protein farnesyltransferase in plants. Molecular cloning and expression of a homolog of the beta subunit from the garden pea.

Z Yang 1, C L Cramer 1, J C Watson 1
PMCID: PMC160617  PMID: 8278509

Abstract

Protein farnesyltransferase is a heterodimeric enzyme that attaches a farnesyl moiety to C-terminal cysteine residues. Both the alpha and beta subunits have recently been cloned and sequenced from yeast and rat. Degenerate oligonucleotides, corresponding to conserved regions of the beta subunit, were used as primers for the polymerase chain reaction to amplify cDNA synthesized from total cellular RNA from the apical buds of pea (Pisum sativum L.) seedlings. The 171-bp fragment obtained encodes an open reading frame of 57 amino acids showing 65% identity to the rat protein farnesyltransferase beta subunit. Using this fragment to screen a pea cDNA library, one full-length cDNA clone, designated PsFTb, was obtained that contains an open reading frame encoding a polypeptide of 419 amino acids. The predicted amino acid sequence exhibits 48 and 40% identity to the rat and yeast beta subunits, respectively, indicating that this cDNA encodes a pea homolog of the beta subunit of farnesyltransferase. Gel blot hybridizations show that PsFTb is likely to be encoded by a single-copy gene and is expressed as a transcript of approximately 1.7 kb. During photoregulated leaf development in continuous white light, PsFTb transcript levels within apical buds decline by approximately 5-fold.

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Selected References

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  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. Casey P. J., Solski P. A., Der C. J., Buss J. E. p21ras is modified by a farnesyl isoprenoid. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8323–8327. doi: 10.1073/pnas.86.21.8323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Casey P. J., Thissen J. A., Moomaw J. F. Enzymatic modification of proteins with a geranylgeranyl isoprenoid. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8631–8635. doi: 10.1073/pnas.88.19.8631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen W. J., Andres D. A., Goldstein J. L., Brown M. S. Cloning and expression of a cDNA encoding the alpha subunit of rat p21ras protein farnesyltransferase. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11368–11372. doi: 10.1073/pnas.88.24.11368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Fenton R. G., Kung H. F., Longo D. L., Smith M. R. Regulation of intracellular actin polymerization by prenylated cellular proteins. J Cell Biol. 1992 Apr;117(2):347–356. doi: 10.1083/jcb.117.2.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Finegold A. A., Johnson D. I., Farnsworth C. C., Gelb M. H., Judd S. R., Glomset J. A., Tamanoi F. Protein geranylgeranyltransferase of Saccharomyces cerevisiae is specific for Cys-Xaa-Xaa-Leu motif proteins and requires the CDC43 gene product but not the DPR1 gene product. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4448–4452. doi: 10.1073/pnas.88.10.4448. [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. Gibbs J. B. Ras C-terminal processing enzymes--new drug targets? Cell. 1991 Apr 5;65(1):1–4. doi: 10.1016/0092-8674(91)90352-y. [DOI] [PubMed] [Google Scholar]
  11. Glomset J. A., Gelb M. H., Farnsworth C. C. Prenyl proteins in eukaryotic cells: a new type of membrane anchor. Trends Biochem Sci. 1990 Apr;15(4):139–142. doi: 10.1016/0968-0004(90)90213-u. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Goodman L. E., Perou C. M., Fujiyama A., Tamanoi F. Structure and expression of yeast DPR1, a gene essential for the processing and intracellular localization of ras proteins. Yeast. 1988 Dec;4(4):271–281. doi: 10.1002/yea.320040405. [DOI] [PubMed] [Google Scholar]
  14. Hall A. The cellular functions of small GTP-binding proteins. Science. 1990 Aug 10;249(4969):635–640. doi: 10.1126/science.2116664. [DOI] [PubMed] [Google Scholar]
  15. Hancock J. F., Cadwallader K., Paterson H., Marshall C. J. A CAAX or a CAAL motif and a second signal are sufficient for plasma membrane targeting of ras proteins. EMBO J. 1991 Dec;10(13):4033–4039. doi: 10.1002/j.1460-2075.1991.tb04979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hancock J. F., Magee A. I., Childs J. E., Marshall C. J. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell. 1989 Jun 30;57(7):1167–1177. doi: 10.1016/0092-8674(89)90054-8. [DOI] [PubMed] [Google Scholar]
  17. Inglese J., Glickman J. F., Lorenz W., Caron M. G., Lefkowitz R. J. Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase. J Biol Chem. 1992 Jan 25;267(3):1422–1425. [PubMed] [Google Scholar]
  18. Johnson D. I., O'Brien J. M., Jacobs C. W. Isolation and sequence analysis of CDC43, a gene involved in the control of cell polarity in Saccharomyces cerevisiae. Gene. 1990 May 31;90(1):93–98. doi: 10.1016/0378-1119(90)90443-u. [DOI] [PubMed] [Google Scholar]
  19. Kawata M., Farnsworth C. C., Yoshida Y., Gelb M. H., Glomset J. A., Takai Y. Posttranslationally processed structure of the human platelet protein smg p21B: evidence for geranylgeranylation and carboxyl methylation of the C-terminal cysteine. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8960–8964. doi: 10.1073/pnas.87.22.8960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kohl N. E., Diehl R. E., Schaber M. D., Rands E., Soderman D. D., He B., Moores S. L., Pompliano D. L., Ferro-Novick S., Powers S. Structural homology among mammalian and Saccharomyces cerevisiae isoprenyl-protein transferases. J Biol Chem. 1991 Oct 5;266(28):18884–18888. [PubMed] [Google Scholar]
  21. Krohne G., Waizenegger I., Höger T. H. The conserved carboxy-terminal cysteine of nuclear lamins is essential for lamin association with the nuclear envelope. J Cell Biol. 1989 Nov;109(5):2003–2011. doi: 10.1083/jcb.109.5.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin X., Feng X. H., Watson J. C. Differential accumulation of transcripts encoding protein kinase homologs in greening pea seedlings. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6951–6955. doi: 10.1073/pnas.88.16.6951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lütcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. Selection of AUG initiation codons differs in plants and animals. EMBO J. 1987 Jan;6(1):43–48. doi: 10.1002/j.1460-2075.1987.tb04716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maltese W. A. Posttranslational modification of proteins by isoprenoids in mammalian cells. FASEB J. 1990 Dec;4(15):3319–3328. doi: 10.1096/fasebj.4.15.2123808. [DOI] [PubMed] [Google Scholar]
  25. Matsui M., Sasamoto S., Kunieda T., Nomura N., Ishizaki R. Cloning of ara, a putative Arabidopsis thaliana gene homologous to the ras-related gene family. Gene. 1989;76(2):313–319. doi: 10.1016/0378-1119(89)90171-6. [DOI] [PubMed] [Google Scholar]
  26. McElrath M. J., Kaplan G., Burkhardt R. A., Cohn Z. A. Cutaneous response to recombinant interleukin 2 in human immunodeficiency virus 1-seropositive individuals. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5783–5787. doi: 10.1073/pnas.87.15.5783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Narita J. O., Gruissem W. Tomato hydroxymethylglutaryl-CoA reductase is required early in fruit development but not during ripening. Plant Cell. 1989 Feb;1(2):181–190. doi: 10.1105/tpc.1.2.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ohya Y., Goebl M., Goodman L. E., Petersen-Bjørn S., Friesen J. D., Tamanoi F., Anraku Y. Yeast CAL1 is a structural and functional homologue to the DPR1 (RAM) gene involved in ras processing. J Biol Chem. 1991 Jul 5;266(19):12356–12360. [PubMed] [Google Scholar]
  29. Palme K., Diefenthal T., Vingron M., Sander C., Schell J. Molecular cloning and structural analysis of genes from Zea mays (L.) coding for members of the ras-related ypt gene family. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):787–791. doi: 10.1073/pnas.89.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Reiss Y., Seabra M. C., Armstrong S. A., Slaughter C. A., Goldstein J. L., Brown M. S. Nonidentical subunits of p21H-ras farnesyltransferase. Peptide binding and farnesyl pyrophosphate carrier functions. J Biol Chem. 1991 Jun 5;266(16):10672–10677. [PubMed] [Google Scholar]
  31. Reiss Y., Stradley S. J., Gierasch L. M., Brown M. S., Goldstein J. L. Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):732–736. doi: 10.1073/pnas.88.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schafer W. R., Trueblood C. E., Yang C. C., Mayer M. P., Rosenberg S., Poulter C. D., Kim S. H., Rine J. Enzymatic coupling of cholesterol intermediates to a mating pheromone precursor and to the ras protein. Science. 1990 Sep 7;249(4973):1133–1139. doi: 10.1126/science.2204115. [DOI] [PubMed] [Google Scholar]
  33. Schmidt R. A., Schneider C. J., Glomset J. A. Evidence for post-translational incorporation of a product of mevalonic acid into Swiss 3T3 cell proteins. J Biol Chem. 1984 Aug 25;259(16):10175–10180. [PubMed] [Google Scholar]
  34. Seabra M. C., Reiss Y., Casey P. J., Brown M. S., Goldstein J. L. Protein farnesyltransferase and geranylgeranyltransferase share a common alpha subunit. Cell. 1991 May 3;65(3):429–434. doi: 10.1016/0092-8674(91)90460-g. [DOI] [PubMed] [Google Scholar]
  35. Terryn N., Anuntalabhochai S., Van Montagu M., Inzé D. Analysis of a Nicotiana plumbaginifolia cDNA encoding a novel small GTP-binding protein. FEBS Lett. 1992 Mar 16;299(3):287–290. doi: 10.1016/0014-5793(92)80133-2. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Yamane H. K., Farnsworth C. C., Xie H. Y., Evans T., Howald W. N., Gelb M. H., Glomset J. A., Clarke S., Fung B. K. Membrane-binding domain of the small G protein G25K contains an S-(all-trans-geranylgeranyl)cysteine methyl ester at its carboxyl terminus. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):286–290. doi: 10.1073/pnas.88.1.286. [DOI] [PMC free article] [PubMed] [Google Scholar]

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