Skip to main content
SpringerLink
Log in

Distinct domains of human CDC5 direct its nuclear import and association with the spliceosome

  • Original Article
  • Published:
Cell Biochemistry and Biophysics Aims and scope Submit manuscript

Abstract

Genetic studies have shown that CDC5 proteins are essential for G2 progression and mitotic entry. CDC5 homologs in yeast and mammals are essential for pre-messenger ribonucleic acid (mRNA) processing. Other gene products also have been shown to play roles in both pre-mRNA splicing and cell cycle regulation, prompting the description of several models to explain the mechanism(s) linking these two processes. In this study, we demonstrate that the amino-terminus of human CDC5 directs nuclear import independent of consensus nuclear localization signals or phosphorylation, and that the carboxyl-terminus of human CDC5 preferentially associates with spliceosomal complexes in proximity of RNA transcription during interphase. hCDC5 colocalizes with Sm proteins in a cell cycle- and domain-dependent manner, and several proteins in the human CDC5-associated complex are identified that suggest potential roles for the complex in coupling pre-mRNA splicing to transcriptional activation and protein translation. These results indicate that human CDC5 may function in pre-mRNA processing and cell cycle progression through more than one mechanism.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Nasmyth, K. and Nurse, P. (1981) Cell division cycle mutants altered in DNA replication and mitosis in the fission yeast Schizosaccharomyces pombe. Mol. Gen. Genet. 182, 119–124.

    Article  PubMed  CAS  Google Scholar 

  2. Ohi, R., McCollum, D., Hirani, B., Den Haese, G. J., Zhang, X., Burke, J. D., Turner, K. and Gould, K. L. (1994) The Schizosaccharomyces pombe cdc5+gene encodes an essential protein with homology to c-Myb. EMBO J. 13, 471–483.

    PubMed  CAS  Google Scholar 

  3. Bernstein, H. S. and Coughlin, S. R. (1998) A mammalian homolog of fission yeast Cdc5 regulates G2 progression and mitotic entry. J. Biol. Chem. 273, 4666–4671.

    Article  PubMed  CAS  Google Scholar 

  4. Bernstein, H. S. and Coughlin, S. R. (1997) Pombe Cdc5-related protein. A putative human transcription factor implicated in mitogen-activated signaling. J. Biol. Chem. 272, 5833–5837.

    Article  PubMed  CAS  Google Scholar 

  5. Lei, X.-H., Shen, X., Xu, X.-Q. and Bernstein, H. S. (2000) Human Cdc5, a regulator of mitotic entry, can act as a site-specific DNA binding protein. J. Cell. Sci. 113, 4523–4531.

    PubMed  CAS  Google Scholar 

  6. Johnson, L. M. and Too, C. K. (2001) Prolactin, interleukin-2 and FGF-2 stimulate expression, nuclear distribution and DNA-binding of rat homolog of pombe Cdc5 in Nb2 T lymphoma cells. Mol. Cell. Endocrinol. 184, 151–161.

    Article  PubMed  CAS  Google Scholar 

  7. Tsai, W. Y., Chow, Y. T., Chen, H. R., Huang, K. T., Hong, R. I., Jan, S. P., et al. (1999) Cef1p is a component of the Prp 19p-associated complex and essential for pre-mRNA splicing. J. Biol. Chem. 274, 9455–9462.

    Article  PubMed  CAS  Google Scholar 

  8. McDonald, W. H., Ohi, R., Smelkova, N., Frendewey, D. and Gould, K. L. (1999) Mybrelated fission yeast cdc5p is a component of a 40S snRNP-containing complex and is essential for pre-mRNA splicing. Mol. Cell. Biol. 19, 5352–5362.

    PubMed  CAS  Google Scholar 

  9. Ajuh, P., Kuster, B., Panov, K., Zomerdijk, J. C., Mann, M. and Lamond, A. I. (2000) Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry. EMBO. J. 19, 6569–6581.

    Article  PubMed  CAS  Google Scholar 

  10. Ohi, M. D., Link, A. J., Ren, L., Jennings, J. L., McDonald, W. H. and Gould, K. L. (2002) Proteomics analysis reveals stable multiprotein complexes in both fission and budding yeasts containing Myb-related Cdc5p/Cef1p, novel pre-mRNA splicing factors, and snRNAs. Mol. Cell. Biol. 22, 2011–2024.

    Article  PubMed  CAS  Google Scholar 

  11. Zhou, Z., Licklider, L. J., Gygi, S. P. and Reed, R. (2002) comprehensive proteomic analysis of the human spliceosome. Nature 419, 182–185.

    Article  PubMed  CAS  Google Scholar 

  12. Makarov, E. M., Makarova, O. V., Urlaub, H., Gentzel, M., Will, C. L., Wilm, M. and Luhrmann, R. (2002) Small nuclear ribonucleo-protein remodeling during catalytic activation of the spliceosome. Science 298, 2205–2208.

    Article  PubMed  CAS  Google Scholar 

  13. Burns, C. G., Ohi, R., Krainer, A. R. and Gould, K. L. (1999) Evidence that Myb-related CDC5 proteins are required for pre-mRNA splicing. Proc. Natl. Acad. Sci. USA 96, 13789–13794.

    Article  PubMed  CAS  Google Scholar 

  14. Boudrez, A., Beullens, M., Groenen, P., Van Eynde, A., Vulsteke, V., Jagiello, I., et al. (2000) NIPP1-mediated interaction of protein phosphatase-1 with CDC5L, a regulator of pre-mRNA splicing and mitotic entry. J. Biol. Chem. 275, 25411–25417.

    Article  PubMed  CAS  Google Scholar 

  15. Ajuh, P., Sleeman, J., Chusainow, J. and Lamond, A. I. (2001) A direct interaction between the carboxyl-terminal region of CDC5L and the WD40 domain of PLRG1 is essential for pre-mRNA splicing. J. Biol. Chem. 276, 42370–42381.

    Article  PubMed  CAS  Google Scholar 

  16. Burns, C. G., Ohi, R., Mehta, S., O'Toole, E. T., Winey, M., Clark, et al. (2002) Removal of a single alpha-tubulin gene intron suppresses cell cycle arrest phenotypes of splicing factor mutations in Saccharomyces cerevisiae. Mol. Cell. Biol. 22, 801–815.

    Article  PubMed  CAS  Google Scholar 

  17. Morgan, D. O. (1997) Cyclin-dependent kinases: engines, clocks, and microprocessors. Ann. Rev. Cell Dev. Biol. 13, 261–291.

    Article  CAS  Google Scholar 

  18. Johnston, L. H. and Thomas, A. P. (1982) The isolation of new DNA synthesis mutants in the yeast Saccharomyces cerevisiae. Mol. Gen. Genet. 186, 439–444.

    Article  PubMed  CAS  Google Scholar 

  19. Vijayraghavan, U., Company, M. and Abelson, J. (1989) Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae. Genes Dev. 3, 1206–1216.

    Article  PubMed  CAS  Google Scholar 

  20. Shea, J. E., Toyn, J. H. and Johnston, L. H. (1994) The budding yeast U5 snRNP Prp8 is a highly conserved protein which links RNA splicing with cell cycle progression. Nuclo Acids Res. 22, 5555–5564.

    Article  CAS  Google Scholar 

  21. Kassir, Y. and Simchen, G. (1978) Meiotic recombination and DNA synthesis in a new cell cycle mutant of Saccharomyces cerevisiae. Genetics 90, 49–68.

    PubMed  CAS  Google Scholar 

  22. Kassir, Y., Kupiec, M., Shalom, A. and Simchen, G. (1985) Cloning and mapping of CDC40, a Saccharomyces cerevisiae gene with a role in DNA repair. Curr. Genet. 9, 253–257.

    Article  PubMed  CAS  Google Scholar 

  23. Kupiec, M. and Simchen, G. (1986) DNA-repair characterization of cdc40-1, a cell-cycle mutant of Saccharomyces cerevisiae. Mutat. Res. 162, 33–40.

    PubMed  CAS  Google Scholar 

  24. Vaisman, N., Tsouladze, A., Robzyk, K., Ben-Yehuda, S., Kupiec, M. and Kassir, Y. (1995) The role of Saccharomyces cerevisiae Cdc40p in DNA replication and mitotic spindle formation and/or maintenance. Mol. Gen. Genet. 247, 123–136.

    Article  PubMed  CAS  Google Scholar 

  25. Hwang, L. H. and Murray, A. W. (1997) A novel yeast screen for mitotic arrest mutants identifies DOC1, a new gene involved in cyclin proteolysis. Mol. Biol. Cell. 8, 1877–1887.

    PubMed  CAS  Google Scholar 

  26. Company, M., Arenas, J. and Abelson, J. (1991) Requirement of the RNA helicase-like protein PRP22 for release of messenger RNA from spliceosomes. Nature 349, 487–493.

    Article  PubMed  CAS  Google Scholar 

  27. Lundgren, K., Allan, S., Urushiyama, S., Tani, T., Ohshima, Y., Frendewey, D. and Beach, D. (1996) A connection between pre-mRNA splicing and the cell cycle in fission yeast: cdc28+ is allelic with prp8+ and encodes an RNA-dependent ATPase/helicase. Mol. Biol. Cell. 7, 1083–1094.

    PubMed  CAS  Google Scholar 

  28. Potashkin, J., Naik, K. and Wentz-Hunter, K. (1993) U2AF homolog required for splicing in vivo. Science 262, 573–575.

    Article  PubMed  CAS  Google Scholar 

  29. Takahashi, K., Yamada, H. and Yanagida, M. (1994) Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality. Mol. Biol. Cell. 5, 1145–1158.

    PubMed  CAS  Google Scholar 

  30. Gross, T., Richert, K., Mierke, C., Lutzelberger, M. and Kaufer, N. F. (1998) Identification and characterization of srpl, a gene of fission yeast encoding a RNA binding domain and a RS domain typical of SR splicing factors. Nucl. Acids Res. 26, 505–511.

    Article  PubMed  CAS  Google Scholar 

  31. Rosenberg, G. H., Alahari, S. K. and Kaufer, N. F. (1991) prp4 from Schizosaccharomyces pombe, a mutant deficient in pre-mRNA splicing isolated using genes containing artificial introns. Mol. Gen. Genet. 226, 305–309.

    Article  PubMed  CAS  Google Scholar 

  32. Tang, Z., Yanagida, M. and Lin, R. J. (1998) Fission yeast mitotic regulator Dsk1 is an SR protein-specific kinase. J. Biol. Chem. 273, 5963–5969.

    Article  PubMed  CAS  Google Scholar 

  33. Spector, D. L. (1994) RNA processing. Cycling splicing factors. Nature 369, 604.

    Article  PubMed  CAS  Google Scholar 

  34. Seshadri, V., Vaidya, V. C. and Vijayraghavan, U. (1996) Genetic studies of the PRP17 gene of Saccharomyces cerevisiae: A domain essential for function maps to a nonconserved region of the protein. Genetics 143, 45–55.

    PubMed  CAS  Google Scholar 

  35. Stukenberg, P. T., Lustig, K. D., McGarry, T. J., King, R. W., Kuang, J. and Kirschner, M. W. (1997) Systematic identification of mitotic phosphoproteins. Curr. Biol. 7, 338–348.

    Article  PubMed  CAS  Google Scholar 

  36. Mintz, P. J. and Spector, D. L. (2000) Compartmentalization of RNA processing factors within nuclear speckles. J. Struct. Biol. 129, 241–251.

    Article  PubMed  CAS  Google Scholar 

  37. Dignam, J. D., Lebovitz, R. M. and Roeder, R. G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucl. Acids Res. 11, 1475–1489.

    Article  PubMed  CAS  Google Scholar 

  38. Markovtsov, V., Nikolic, J. M., Goldman, J. A., Turck, C. W., Chou, M. Y. and Black, D. L. (2000) Cooperative assembly of an hnRNP complex induced by a tissue-specific homolog of polypyrimidine tract binding protein. Mol. Cell. Biol. 20, 7463–7479.

    Article  PubMed  CAS  Google Scholar 

  39. Duverger, E., Pellerin-Mendes, C., Mayer, R., Roche, A. C. and Monsigny, M. (1995) Nuclear import of glycoconjugates is distinct from the classical NLS pathway. J. Cell. Sci. 108, 1325–1332.

    PubMed  CAS  Google Scholar 

  40. Shevchenko, A., Wilm, M., Vorm, O. and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850–858.

    Article  PubMed  CAS  Google Scholar 

  41. Eilbracht, J. and Schmidt-Zachmann, M. S. (2001) Identification of a sequence element directing a protein to nuclear speckles. Proc. Natl. Acad. Sci. USA 98, 3849–3854.

    Article  PubMed  CAS  Google Scholar 

  42. Wei, X., Somanathan, S., Samarabandu, J. and Berezney, R. (1999) Three-dimensional visualization of transcription sites and their association with splicing factor-rich nuclear speckles. J. Cell. Biol. 146, 543–558.

    Article  PubMed  CAS  Google Scholar 

  43. Ohi, R., Reoktistova, A., McCann, S., Valentine, V., Look, A. T., Lipsick, J. S. and Gould, K. L. (1998) Myb-related Schizosaccharomyces pombe cdc5p is structurally and functionally conserved in eukaryotes. Mol. Cell. Biol. 18, 4097–4108.

    PubMed  CAS  Google Scholar 

  44. van der Houven van Oordt, W., Diaz-Meco, M. T., Lozano, J., Krainer, A. R., Moscat, J. and Caceres, J. F. (2000) The MKK(3/6)-p38-signaling cascade alters the subcellular distribution of hnRNP A1 and modulates alternative splicing regulation. J. Cell. Biol. 149, 307–316.

    Article  PubMed  Google Scholar 

  45. Tollervey, D. and Caceres, J. F. (2000) RNA processing marches on. Cell 103, 703–709.

    Article  PubMed  CAS  Google Scholar 

  46. Engemann, H., Heinzel, V., Page, G., Preuss, U. and Scheidtmann, K. H. (2002) DAP-like kinase interacts with the rat homolog of Schizosaccharomyces pombe CDC5 protein, a factor involved in pre-mRNA splicing and required for G2/M phase transition. Nucl. Acids Res. 30, 1408–1417.

    Article  PubMed  CAS  Google Scholar 

  47. Zhang, D., Abovich, N. and Rosbash, M. (2001) A biochemical function for the Sm complex. Mol. Cell. 7, 319–329.

    Article  PubMed  CAS  Google Scholar 

  48. Venables, J. P., Elliott, D. J., Makarova, O. V., Makarov, E. M., Cooke, H. J. and Eperon, I. C. (2000) RBMY, a probable human spermatogenesis factor, and other hnRNP G proteins interact with Tra2beta and affect splicing. Hum. Mol. Genet. 9, 685–694.

    Article  PubMed  CAS  Google Scholar 

  49. Gotzmann, J., Gerner, C., Meissner, M., Holzmann, K., Grimm, R., Mikulits, W. and Sauermann, G. (2000) hNMP 200: a novel human common nuclear matrix protein combining structural and regulatory functions. Exp. Cell. Res. 261, 166–179.

    Article  PubMed  CAS  Google Scholar 

  50. Sasaki, Y., Itoh, F., Suzuki, H., Kobayashi, T., Kakiuchi, H., Hareyama, M. and Imai, K. (2000) Identification of genes highly expressed in G2-arrested Chinese hamster ovary cells by differential display analysis. J. Clin. Lab. Anal. 14, 314–319.

    Article  PubMed  CAS  Google Scholar 

  51. Serfling, E., Berberich-Siebelt, F., Chuvpilo, S., Jankevics, E., Klein-Hessling, S., Twardzik, T. and Avots, A. (2000) The role of NF-AT transcription factors in T cell activation and differentiation. Biochim. Biophys. Acta 1498, 1–18.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cardiovascular Research Institute, University of California San Francisco, 505 Parnassus Avenue, Box 0130, 94143-0130, San Francisco, CA

    Liansen Liu, Remo Gräub, Myint Hlaing, Conrad L. Epting, Leanne Zhang & Harold S. Bernstein MD, PhD

  2. Howard Hughes Medical Institute, University of California San Francisco, 505 Parnassus Avenue, Box 0130, 94143-0130, San Francisco, CA

    Christoph W. Turck

  3. Cancer Center, University of California San Francisco, 505 Parnassus Avenue, Box 0130, 94143-0130, San Francisco, CA

    Harold S. Bernstein MD, PhD

  4. Department of Pediatrics, University of California San Francisco, 505 Parnassus Avenue, Box 0130, 94143-0130, San Francisco, CA

    Conrad L. Epting, Xiao-Qin Xu & Harold S. Bernstein MD, PhD

Authors
  1. Liansen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Remo Gräub
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Myint Hlaing
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Conrad L. Epting
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Christoph W. Turck
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiao-Qin Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Leanne Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Harold S. Bernstein MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Harold S. Bernstein MD, PhD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, L., Gräub, R., Hlaing, M. et al. Distinct domains of human CDC5 direct its nuclear import and association with the spliceosome. Cell Biochem Biophys 39, 119–131 (2003). https://doi.org/10.1385/CBB:39:2:119

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1385/CBB:39:2:119

Index Entries

Search

Navigation