Home LiteratureArticle Details
PMID: 1722313 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Senescent cells fail to express cdc2, cycA, and cycB in response to mitogen stimulation.

Stein GH, Drullinger LF, Robetorye RS, Pereira-Smith OM, Smith JR

Abstract

Senescent human diploid fibroblasts (HDF) contain no detectable cdc2 mRNA or p34cdc2 protein. Similarly, young quiescent HDF have only low levels of cdc2 mRNA and protein. After serum stimulation, quiescent HDF accumulate increasing amounts of cdc2 mRNA and protein and go through DNA synthesis and mitosis. In contrast, serum-stimulated senescent HDF fail to accumulate detectable amounts of cdc2 mRNA and protein and fail to enter S phase. Mitosis is likewise deficient in senescent cells even when they have been induced to synthesize DNA by simian virus 40 large tumor antigen. Since p34cdc2 or its homologues appear to be required for DNA synthesis and mitosis in eukaryotes, a lack of these molecules in serum-stimulated senescent HDF could be an important reason for their inability to enter S phase or mitosis. Nuclear microinjection of cdc2 DNA into senescent HDF causes rounding up of the cells but no induction of DNA synthesis. Since cyclins A and B are important cofactors of the protein kinase activity of p34cdc2 or its homologues, we analyzed expression of these genes in serum-stimulated senescent HDF and determined that they contain little or no cycA or cycB mRNA. These deficiencies may be relevant to the lack of DNA synthesis and mitosis in senescent HDF.

Related Genes
MeSH Terms
Amino Acid Sequence Blotting, Northern CDC2 Protein Kinase/genetics Cell Line Cell Nucleus/physiology Cellular Senescence DNA Probes DNA Replication Fibroblasts/cytology,physiology Gene Expression Humans Immunoblotting Mitogens/pharmacology Mitosis Molecular Sequence Data Poly A/genetics,isolation & purification RNA/genetics,isolation & purification RNA, Messenger/genetics Transcription, Genetic Transfection
Chemicals
DNA Probes Mitogens RNA, Messenger Poly A RNA CDC2 Protein Kinase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Stein G H
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309-0347.
Drullinger L F
Robetorye R S
Pereira-Smith O M
Smith J R
References (33)
33 references, click to expand
  1. The FT210 cell line is a mouse G2 phase mutant with a temperature-sensitive CDC2 gene product.
    Cell. 1990 Oct 19;63(2):313-24 PMID: 2208288
  2. Hepatitis B virus integration in a cyclin A gene in a hepatocellular carcinoma.
    Nature. 1990 Feb 8;343(6258):555-7 PMID: 1967822
  3. Identification of p34 and p13, human homologs of the cell cycle regulators of fission yeast encoded by cdc2+ and suc1+.
    Cell. 1987 Jul 17;50(2):319-25 PMID: 3297353
  4. Failure to phosphorylate the retinoblastoma gene product in senescent human fibroblasts.
    Science. 1990 Aug 10;249(4969):666-9 PMID: 2166342
  5. Human cyclin A is adenovirus E1A-associated protein p60 and behaves differently from cyclin B.
    Nature. 1990 Aug 23;346(6286):760-3 PMID: 2143810
  6. Universal control mechanism regulating onset of M-phase.
    Nature. 1990 Apr 5;344(6266):503-8 PMID: 2138713
  7. The retinoblastoma gene and cell growth control.
    Trends Biochem Sci. 1990 May;15(5):199-202 PMID: 2193439
  8. Periodic biosynthesis of the human M-phase promoting factor catalytic subunit p34 during the cell cycle.
    Mol Cell Biol. 1990 Jul;10(7):3847-51 PMID: 2192266
  9. Microinjection of p34cdc2 kinase induces marked changes in cell shape, cytoskeletal organization, and chromatin structure in mammalian fibroblasts.
    Cell. 1990 Jan 12;60(1):151-65 PMID: 2403841
  10. A cellular oncogene (c-Ki-ras) is amplified, overexpressed, and located within karyotypic abnormalities in mouse adrenocortical tumour cells.
    Nature. 1983 Jun 9-15;303(5917):497-501 PMID: 6304530
  11. Complementary DNA sequence of a human cytoplasmic actin. Interspecies divergence of 3' non-coding regions.
    J Mol Biol. 1983 Feb 5;163(4):673-8 PMID: 6842590
  12. Carcinogen-transformed human cells are inhibited from entry into S phase by fusion to senescent cells but cells transformed by DNA tumor viruses overcome the inhibition.
    Proc Natl Acad Sci U S A. 1982 Sep;79(17):5287-91 PMID: 6957863
  13. Dominance of the senescent phenotype in heterokaryons between replicative and post-replicative human fibroblast-like cells.
    Proc Natl Acad Sci U S A. 1974 Jun;71(6):2231-5 PMID: 4366757
  14. A new human p34 protein kinase, CDK2, identified by complementation of a cdc28 mutation in Saccharomyces cerevisiae, is a homolog of Xenopus Eg1.
    EMBO J. 1991 Sep;10(9):2653-9 PMID: 1714386
  15. Isolation of the human cdk2 gene that encodes the cyclin A- and adenovirus E1A-associated p33 kinase.
    Nature. 1991 Sep 12;353(6340):174-7 PMID: 1653904
  16. Evidence that the G1-S and G2-M transitions are controlled by different cdc2 proteins in higher eukaryotes.
    Cell. 1991 Aug 23;66(4):731-42 PMID: 1652371
  17. The cell cycle: spinning farther afield.
    Science. 1991 Jun 14;252(5012):1490-2 PMID: 1828620
  18. Retinoblastoma cancer suppressor gene product is a substrate of the cell cycle regulator cdc2 kinase.
    EMBO J. 1991 Apr;10(4):857-64 PMID: 2009861
  19. Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2.
    Nature. 1987 May 7-13;327(6117):31-5 PMID: 3553962
  20. Synthesis of p34, the mammalian homolog of the yeast cdc2+/CDC28 protein kinase, is stimulated during adenovirus-induced proliferation of primary baby rat kidney cells.
    Oncogene. 1988 Jun;2(6):553-7 PMID: 2838787
  21. Reinitiation of cellular DNA synthesis in BrdU-selected nondividing senescent WI-38 cells by simian virus 40 infection.
    J Cell Physiol. 1985 Oct;125(1):122-6 PMID: 2995423
  22. Entry into S phase is inhibited in human fibroblasts by rat liver poly(A)+RNA.
    Exp Cell Res. 1985 Oct;160(2):544-9 PMID: 2412871
  23. A 60 kd cdc2-associated polypeptide complexes with the E1A proteins in adenovirus-infected cells.
    Cell. 1989 Sep 8;58(5):981-90 PMID: 2570639
  24. Isolation of a human cyclin cDNA: evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2.
    Cell. 1989 Sep 8;58(5):833-46 PMID: 2570636
  25. In vitro phosphorylation of the tumor suppressor gene RB protein by mitosis-specific histone H1 kinase.
    Biochem Biophys Res Commun. 1989 Oct 16;164(1):580-6 PMID: 2553023
  26. The cdc2 kinase is a nuclear protein that is essential for mitosis in mammalian cells.
    Cell. 1989 May 5;57(3):393-401 PMID: 2541912
  27. Regulation of the product of a possible human cell cycle control gene CDC2Hs in B-cells by alpha-interferon and phorbol ester.
    J Biol Chem. 1989 Aug 15;264(23):13697-700 PMID: 2668271
  28. Histone gene expression remains coupled to DNA synthesis during in vitro cellular senescence.
    Exp Cell Res. 1987 Oct;172(2):397-403 PMID: 3653264
  29. Mitogenic effects of the proto-oncogene and oncogene forms of c-H-ras DNA in human diploid fibroblasts.
    Mol Cell Biol. 1986 Aug;6(8):2990-3 PMID: 3785220
  30. Analysis of the G1 arrest position of senescent WI38 cells by quinacrine dihydrochloride nuclear fluorescence. Evidence for a late G1 arrest.
    Exp Cell Res. 1986 Nov;167(1):87-94 PMID: 3758211
  31. Repression of c-fos transcription and an altered genetic program in senescent human fibroblasts.
    Science. 1990 Jan 12;247(4939):205-9 PMID: 2104680
  32. Negative regulation of human c-fos expression by the retinoblastoma gene product.
    Nature. 1990 Aug 16;346(6285):668-71 PMID: 2117257
  33. Regulation of ornithine decarboxylase and other cell cycle-dependent genes during senescence of IMR-90 human diploid fibroblasts.
    J Biol Chem. 1988 Aug 15;263(23):11431-5 PMID: 3403538
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1991-12-15
Pages
11012-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC53063
Subset
IM
Grants
NIA NIH HHS · AG00947 · United States
NIA NIH HHS · AG05333 · United States
NIA NIH HHS · T32-AG00183 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]