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. 1990 Feb 1;110(2):255–261. doi: 10.1083/jcb.110.2.255

Characterization of two populations of statin and the relationship of their syntheses to the state of cell proliferation

PMCID: PMC2116018  PMID: 2298807

Abstract

Statin has previously been identified to be a 57-kD protein present in the nuclei of quiescent and senescent human fibroblasts, but not in their replicating counterparts (Wang, E. 1985. J. Cell Biol. 100: 545- 551). In the present report we demonstrate by immunoprecipitation analysis of fractionated cellular extracts the existence of two populations of statin. The Triton X-100-soluble statin is found in replicating sparse cultures as well as in quiescent confluent cultures and quiescent serum-starved cultures of young human fibroblasts, but the Triton X-100-insoluble, nuclear envelope-localized statin is present only in the quiescent cultures. Two-dimensional gel analysis of the immunoprecipitated cellular fractions reveals that both populations of statin have an isoelectric point of 5.3. Pulse-chase experiments show that statin is synthesized as a 57-kD polypeptide and is not processed from a precursor of different molecular mass. Experiments on serum stimulation of quiescent cells show that synthesis of the Triton X-100-insoluble statin decreases rapidly during the transition from the G0 to S phase, and that this decrease is accompanied by a slower reduction in synthesis of the Triton X-100-soluble statin. These results suggest that the cellular expression of the two populations of statin may be associated with the mechanisms controlling the transition between the growing state and the quiescent state and confirm the previous finding that the Triton X-100-insoluble, nuclear envelope- localized statin could be used as a marker for cells arrested at the G0 phase of the cell cycle.

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Selected References

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  1. Bravo R., Celis J. E. A search for differential polypeptide synthesis throughout the cell cycle of HeLa cells. J Cell Biol. 1980 Mar;84(3):795–802. doi: 10.1083/jcb.84.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bravo R., Fey S. J., Bellatin J., Larsen P. M., Arevalo J., Celis J. E. Identification of a nuclear and of a cytoplasmic polypeptide whose relative proportions are sensitive to changes in the rate of cell proliferation. Exp Cell Res. 1981 Dec;136(2):311–319. doi: 10.1016/0014-4827(81)90009-4. [DOI] [PubMed] [Google Scholar]
  3. Bravo R., Frank R., Blundell P. A., Macdonald-Bravo H. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta. Nature. 1987 Apr 2;326(6112):515–517. doi: 10.1038/326515a0. [DOI] [PubMed] [Google Scholar]
  4. Bravo R., Macdonald-Bravo H. Changes in the nuclear distribution of cyclin (PCNA) but not its synthesis depend on DNA replication. EMBO J. 1985 Mar;4(3):655–661. doi: 10.1002/j.1460-2075.1985.tb03679.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bravo R., Macdonald-Bravo H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol. 1987 Oct;105(4):1549–1554. doi: 10.1083/jcb.105.4.1549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bravo R. Synthesis of the nuclear protein cyclin (PCNA) and its relationship with DNA replication. Exp Cell Res. 1986 Apr;163(2):287–293. doi: 10.1016/0014-4827(86)90059-5. [DOI] [PubMed] [Google Scholar]
  7. Celis J. E., Celis A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Proc Natl Acad Sci U S A. 1985 May;82(10):3262–3266. doi: 10.1073/pnas.82.10.3262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Celis J. E., Madsen P., Nielsen S., Celis A. Nuclear patterns of cyclin (PCNA) antigen distribution subdivide S-phase in cultured cells--some applications of PCNA antibodies. Leuk Res. 1986;10(3):237–249. doi: 10.1016/0145-2126(86)90021-4. [DOI] [PubMed] [Google Scholar]
  9. Hollenberg M. D., Cuatrecasas P. Insulin and epidermal growth factor. Human fibroblast receptors related to deoxyribonucleic acid synthesis and amino acid uptake. J Biol Chem. 1975 May 25;250(10):3845–3853. [PubMed] [Google Scholar]
  10. Kurki P., Vanderlaan M., Dolbeare F., Gray J., Tan E. M. Expression of proliferating cell nuclear antigen (PCNA)/cyclin during the cell cycle. Exp Cell Res. 1986 Sep;166(1):209–219. doi: 10.1016/0014-4827(86)90520-3. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Mathews M. B., Bernstein R. M., Franza B. R., Jr, Garrels J. I. Identity of the proliferating cell nuclear antigen and cyclin. Nature. 1984 May 24;309(5966):374–376. doi: 10.1038/309374a0. [DOI] [PubMed] [Google Scholar]
  13. Miyachi K., Fritzler M. J., Tan E. M. Autoantibody to a nuclear antigen in proliferating cells. J Immunol. 1978 Dec;121(6):2228–2234. [PubMed] [Google Scholar]
  14. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  15. Ottaviano Y., Gerace L. Phosphorylation of the nuclear lamins during interphase and mitosis. J Biol Chem. 1985 Jan 10;260(1):624–632. [PubMed] [Google Scholar]
  16. Prelich G., Kostura M., Marshak D. R., Mathews M. B., Stillman B. The cell-cycle regulated proliferating cell nuclear antigen is required for SV40 DNA replication in vitro. Nature. 1987 Apr 2;326(6112):471–475. doi: 10.1038/326471a0. [DOI] [PubMed] [Google Scholar]
  17. Prelich G., Tan C. K., Kostura M., Mathews M. B., So A. G., Downey K. M., Stillman B. Functional identity of proliferating cell nuclear antigen and a DNA polymerase-delta auxiliary protein. Nature. 1987 Apr 2;326(6112):517–520. doi: 10.1038/326517a0. [DOI] [PubMed] [Google Scholar]
  18. Takasaki Y., Fishwild D., Tan E. M. Characterization of proliferating cell nuclear antigen recognized by autoantibodies in lupus sera. J Exp Med. 1984 Apr 1;159(4):981–992. doi: 10.1084/jem.159.4.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tan C. K., Castillo C., So A. G., Downey K. M. An auxiliary protein for DNA polymerase-delta from fetal calf thymus. J Biol Chem. 1986 Sep 15;261(26):12310–12316. [PubMed] [Google Scholar]
  20. Wang E. A 57,000-mol-wt protein uniquely present in nonproliferating cells and senescent human fibroblasts. J Cell Biol. 1985 Feb;100(2):545–551. doi: 10.1083/jcb.100.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wang E. Are cross-bridging structures involved in the bundle formation of intermediate filaments and the decrease in locomotion that accompany cell aging? J Cell Biol. 1985 May;100(5):1466–1473. doi: 10.1083/jcb.100.5.1466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wang E., Gundersen D. Increased organization of cytoskeleton accompanying the aging of human fibroblasts in vitro. Exp Cell Res. 1984 Sep;154(1):191–202. doi: 10.1016/0014-4827(84)90679-7. [DOI] [PubMed] [Google Scholar]
  23. Wang E., Krueger J. G. Application of a unique monoclonal antibody as a marker for nonproliferating subpopulations of cells of some tissue. J Histochem Cytochem. 1985 Jun;33(6):587–594. doi: 10.1177/33.6.3889143. [DOI] [PubMed] [Google Scholar]
  24. Wang E., Lin S. L. Disappearance of statin, a protein marker for non-proliferating and senescent cells, following serum-stimulated cell cycle entry. Exp Cell Res. 1986 Nov;167(1):135–143. doi: 10.1016/0014-4827(86)90211-9. [DOI] [PubMed] [Google Scholar]
  25. Wang E. Rapid disappearance of statin, a nonproliferating and senescent cell-specific protein, upon reentering the process of cell cycling. J Cell Biol. 1985 Nov;101(5 Pt 1):1695–1701. doi: 10.1083/jcb.101.5.1695. [DOI] [PMC free article] [PubMed] [Google Scholar]

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