Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Aug 2;126(4):853–862. doi: 10.1083/jcb.126.4.853

Yeast NPI46 encodes a novel prolyl cis-trans isomerase that is located in the nucleolus

PMCID: PMC2120118  PMID: 8051210

Abstract

We have identified a gene (NPI46) encoding a new prolyl cis-trans isomerase within the nucleolus of the yeast Saccharomyces cerevisiae. The protein encoded by NPI46 was originally found by us in a search for proteins that recognize nuclear localization sequences (NLSs) in vitro. Thus, NPI46 binds to affinity columns that contain a wild-type histone H2B NLS but not a mutant H2B NLS that is incompetent for nuclear localization in vivo. NPI46 has two domains, a highly charged NH2 terminus similar to two other mammalian nucleolar proteins, nucleolin and Nopp140, and a COOH terminus with 45% homology to a family of mammalian and yeast proline isomerases. NPI46 is capable of catalyzing the prolyl cis-trans isomerization of two small synthetic peptides, succinyl-Ala-Leu-Pro-Phe-p-nitroanilide and succinyl-Ala-Ala-Pro-Phe-p- nitroanilide, as measured by a chymotrypsin-coupled spectrophotometric assay. By indirect immunofluorescence we have shown that NPI46 is a nucleolar protein. NPI46 is not essential for cell viability.

Full Text

The Full Text of this article is available as a PDF (1.7 MB).

Selected References

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

  1. Anfinsen C. B. Principles that govern the folding of protein chains. Science. 1973 Jul 20;181(4096):223–230. doi: 10.1126/science.181.4096.223. [DOI] [PubMed] [Google Scholar]
  2. Brandts J. F., Halvorson H. R., Brennan M. Consideration of the Possibility that the slow step in protein denaturation reactions is due to cis-trans isomerism of proline residues. Biochemistry. 1975 Nov 4;14(22):4953–4963. doi: 10.1021/bi00693a026. [DOI] [PubMed] [Google Scholar]
  3. Créancier L., Prats H., Zanibellato C., Amalric F., Bugler B. Determination of the functional domains involved in nucleolar targeting of nucleolin. Mol Biol Cell. 1993 Dec;4(12):1239–1250. doi: 10.1091/mbc.4.12.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Field J., Nikawa J., Broek D., MacDonald B., Rodgers L., Wilson I. A., Lerner R. A., Wigler M. Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method. Mol Cell Biol. 1988 May;8(5):2159–2165. doi: 10.1128/mcb.8.5.2159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Flanagan W. M., Corthésy B., Bram R. J., Crabtree G. R. Nuclear association of a T-cell transcription factor blocked by FK-506 and cyclosporin A. Nature. 1991 Aug 29;352(6338):803–807. doi: 10.1038/352803a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Galat A., Lane W. S., Standaert R. F., Schreiber S. L. A rapamycin-selective 25-kDa immunophilin. Biochemistry. 1992 Mar 3;31(8):2427–2434. doi: 10.1021/bi00123a031. [DOI] [PubMed] [Google Scholar]
  8. Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
  9. Hayano T., Takahashi N., Kato S., Maki N., Suzuki M. Two distinct forms of peptidylprolyl-cis-trans-isomerase are expressed separately in periplasmic and cytoplasmic compartments of Escherichia coli cells. Biochemistry. 1991 Mar 26;30(12):3041–3048. doi: 10.1021/bi00226a009. [DOI] [PubMed] [Google Scholar]
  10. Heitman J., Movva N. R., Hall M. N. Proline isomerases at the crossroads of protein folding, signal transduction, and immunosuppression. New Biol. 1992 May;4(5):448–460. [PubMed] [Google Scholar]
  11. Heitman J., Movva N. R., Hall M. N. Targets for cell cycle arrest by the immunosuppressant rapamycin in yeast. Science. 1991 Aug 23;253(5022):905–909. doi: 10.1126/science.1715094. [DOI] [PubMed] [Google Scholar]
  12. Herrera A. H., Olson M. O. Association of protein C23 with rapidly labeled nucleolar RNA. Biochemistry. 1986 Oct 7;25(20):6258–6264. doi: 10.1021/bi00368a063. [DOI] [PubMed] [Google Scholar]
  13. Hoffman D. W., Query C. C., Golden B. L., White S. W., Keene J. D. RNA-binding domain of the A protein component of the U1 small nuclear ribonucleoprotein analyzed by NMR spectroscopy is structurally similar to ribosomal proteins. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2495–2499. doi: 10.1073/pnas.88.6.2495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jin Y. J., Burakoff S. J. The 25-kDa FK506-binding protein is localized in the nucleus and associates with casein kinase II and nucleolin. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7769–7773. doi: 10.1073/pnas.90.16.7769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koser P. L., Bergsma D. J., Cafferkey R., Eng W. K., McLaughlin M. M., Ferrara A., Silverman C., Kasyan K., Bossard M. J., Johnson R. K. The CYP2 gene of Saccharomyces cerevisiae encodes a cyclosporin A-sensitive peptidyl-prolyl cis-trans isomerase with an N-terminal signal sequence. Gene. 1991 Dec 1;108(1):73–80. doi: 10.1016/0378-1119(91)90489-x. [DOI] [PubMed] [Google Scholar]
  17. Lapeyre B., Bourbon H., Amalric F. Nucleolin, the major nucleolar protein of growing eukaryotic cells: an unusual protein structure revealed by the nucleotide sequence. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1472–1476. doi: 10.1073/pnas.84.6.1472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee W. C., Xue Z. X., Mélèse T. The NSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs. J Cell Biol. 1991 Apr;113(1):1–12. doi: 10.1083/jcb.113.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Liu J., Walsh C. T. Peptidyl-prolyl cis-trans-isomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4028–4032. doi: 10.1073/pnas.87.11.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Megraw T. L., Chae C. B. Functional complementarity between the HMG1-like yeast mitochondrial histone HM and the bacterial histone-like protein HU. J Biol Chem. 1993 Jun 15;268(17):12758–12763. [PubMed] [Google Scholar]
  21. Meier U. T., Blobel G. Nopp140 shuttles on tracks between nucleolus and cytoplasm. Cell. 1992 Jul 10;70(1):127–138. doi: 10.1016/0092-8674(92)90539-o. [DOI] [PubMed] [Google Scholar]
  22. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Peattie D. A., Harding M. W., Fleming M. A., DeCenzo M. T., Lippke J. A., Livingston D. J., Benasutti M. Expression and characterization of human FKBP52, an immunophilin that associates with the 90-kDa heat shock protein and is a component of steroid receptor complexes. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10974–10978. doi: 10.1073/pnas.89.22.10974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roof W. D., Horne S. M., Young K. D., Young R. slyD, a host gene required for phi X174 lysis, is related to the FK506-binding protein family of peptidyl-prolyl cis-trans-isomerases. J Biol Chem. 1994 Jan 28;269(4):2902–2910. [PubMed] [Google Scholar]
  25. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  26. Sanchez E. R., Faber L. E., Henzel W. J., Pratt W. B. The 56-59-kilodalton protein identified in untransformed steroid receptor complexes is a unique protein that exists in cytosol in a complex with both the 70- and 90-kilodalton heat shock proteins. Biochemistry. 1990 May 29;29(21):5145–5152. doi: 10.1021/bi00473a021. [DOI] [PubMed] [Google Scholar]
  27. Schmid F. X., Grafl R., Wrba A., Beintema J. J. Role of proline peptide bond isomerization in unfolding and refolding of ribonuclease. Proc Natl Acad Sci U S A. 1986 Feb;83(4):872–876. doi: 10.1073/pnas.83.4.872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schmidt-Zachmann M. S., Nigg E. A. Protein localization to the nucleolus: a search for targeting domains in nucleolin. J Cell Sci. 1993 Jul;105(Pt 3):799–806. doi: 10.1242/jcs.105.3.799. [DOI] [PubMed] [Google Scholar]
  29. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Snyder M., Elledge S., Sweetser D., Young R. A., Davis R. W. Lambda gt 11: gene isolation with antibody probes and other applications. Methods Enzymol. 1987;154:107–128. doi: 10.1016/0076-6879(87)54073-3. [DOI] [PubMed] [Google Scholar]
  32. Stamnes M. A., Rutherford S. L., Zuker C. S. Cyclophilins: a new family of proteins involved in intracellular folding. Trends Cell Biol. 1992 Sep;2(9):272–276. doi: 10.1016/0962-8924(92)90200-7. [DOI] [PubMed] [Google Scholar]
  33. Tai P. K., Maeda Y., Nakao K., Wakim N. G., Duhring J. L., Faber L. E. A 59-kilodalton protein associated with progestin, estrogen, androgen, and glucocorticoid receptors. Biochemistry. 1986 Sep 9;25(18):5269–5275. doi: 10.1021/bi00366a043. [DOI] [PubMed] [Google Scholar]
  34. Tropschug M., Barthelmess I. B., Neupert W. Sensitivity to cyclosporin A is mediated by cyclophilin in Neurospora crassa and Saccharomyces cerevisiae. Nature. 1989 Dec 21;342(6252):953–955. doi: 10.1038/342953a0. [DOI] [PubMed] [Google Scholar]
  35. Van Duyne G. D., Standaert R. F., Karplus P. A., Schreiber S. L., Clardy J. Atomic structure of FKBP-FK506, an immunophilin-immunosuppressant complex. Science. 1991 May 10;252(5007):839–842. doi: 10.1126/science.1709302. [DOI] [PubMed] [Google Scholar]
  36. Wülfing C., Lombardero J., Plückthun A. An Escherichia coli protein consisting of a domain homologous to FK506-binding proteins (FKBP) and a new metal binding motif. J Biol Chem. 1994 Jan 28;269(4):2895–2901. [PubMed] [Google Scholar]
  37. Xue Z., Shan X., Lapeyre B., Mélèse T. The amino terminus of mammalian nucleolin specifically recognizes SV40 T-antigen type nuclear localization sequences. Eur J Cell Biol. 1993 Oct;62(1):13–21. [PubMed] [Google Scholar]
  38. Yem A. W., Tomasselli A. G., Heinrikson R. L., Zurcher-Neely H., Ruff V. A., Johnson R. A., Deibel M. R., Jr The Hsp56 component of steroid receptor complexes binds to immobilized FK506 and shows homology to FKBP-12 and FKBP-13. J Biol Chem. 1992 Feb 15;267(5):2868–2871. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES