Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1998 Aug 1;333(Pt 3):645–654. doi: 10.1042/bj3330645

Differential regulation of the retinoblastoma family of proteins during cell proliferation and differentiation.

J Garriga 1, A Limón 1, X Mayol 1, S G Rane 1, J H Albrecht 1, E P Reddy 1, V Andrés 1, X Graña 1
PMCID: PMC1219628  PMID: 9677324

Abstract

In the present study we have analysed the regulation of pocket protein expression and post-transcriptional modifications on cell proliferation and differentiation, both in vivo and in vitro. There are marked changes in pocket protein levels during these transitions, the most striking differences being observed between p130 and p107. The mechanisms responsible for regulating pocket protein levels seem to be dependent on both cell type and pocket protein, in addition to their dependence on the cell growth status. Changes in retinoblastoma protein and p107 levels are independent of their state of phosphorylation. However, whereas p130 phosphorylation to forms characteristic of quiescent/differentiated cells results in the accumulation of p130 protein, phosphorylation of p130 to one or more forms characteristic of cycling cells is accompanied by down-regulation of its protein levels. We also show here that the phosphorylation status and protein levels of p130 and p107 are regulated in vivo as in cultured cells. In vivo, changes in p130 forms are correlated with changes in E2F complexes. Moreover, the modulation of p130 and p107 status during cell differentiation in vitro is consistent with the patterns of protein expression and phosphorylation status found in mouse tissues. Thus in addition to the direct disruption of pocket protein/E2F complexes induced by cyclin/cyclin-dependent kinase, the results we report here indicate that the differential modulation of pocket protein levels constitutes a major mechanism that regulates the pool of each pocket protein that is accessible to E2F and/or other transcription factors.

Full Text

The Full Text of this article is available as a PDF (476.3 KB).

Selected References

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

  1. Albrecht J. H., Hoffman J. S., Kren B. T., Steer C. J. Changes in cell cycle-associated gene expression in a model of impaired liver regeneration. FEBS Lett. 1994 Jun 27;347(2-3):157–162. doi: 10.1016/0014-5793(94)00527-3. [DOI] [PubMed] [Google Scholar]
  2. Albrecht J. H., Hu M. Y., Cerra F. B. Distinct patterns of cyclin D1 regulation in models of liver regeneration and human liver. Biochem Biophys Res Commun. 1995 Apr 17;209(2):648–655. doi: 10.1006/bbrc.1995.1548. [DOI] [PubMed] [Google Scholar]
  3. Andrés V., Walsh K. Myogenin expression, cell cycle withdrawal, and phenotypic differentiation are temporally separable events that precede cell fusion upon myogenesis. J Cell Biol. 1996 Feb;132(4):657–666. doi: 10.1083/jcb.132.4.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beijersbergen R. L., Carlée L., Kerkhoven R. M., Bernards R. Regulation of the retinoblastoma protein-related p107 by G1 cyclin complexes. Genes Dev. 1995 Jun 1;9(11):1340–1353. doi: 10.1101/gad.9.11.1340. [DOI] [PubMed] [Google Scholar]
  5. Clowes A. W., Reidy M. A., Clowes M. M. Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium. Lab Invest. 1983 Sep;49(3):327–333. [PubMed] [Google Scholar]
  6. Cobrinik D., Whyte P., Peeper D. S., Jacks T., Weinberg R. A. Cell cycle-specific association of E2F with the p130 E1A-binding protein. Genes Dev. 1993 Dec;7(12A):2392–2404. doi: 10.1101/gad.7.12a.2392. [DOI] [PubMed] [Google Scholar]
  7. Corbeil H. B., Whyte P., Branton P. E. Characterization of transcription factor E2F complexes during muscle and neuronal differentiation. Oncogene. 1995 Sep 7;11(5):909–920. [PubMed] [Google Scholar]
  8. Fabrikant J. I. The kinetics of cellular proliferation in regenerating liver. J Cell Biol. 1968 Mar;36(3):551–565. doi: 10.1083/jcb.36.3.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fattaey A. R., Harlow E., Helin K. Independent regions of adenovirus E1A are required for binding to and dissociation of E2F-protein complexes. Mol Cell Biol. 1993 Dec;13(12):7267–7277. doi: 10.1128/mcb.13.12.7267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garriga J., Mayol X., Graña X. The CDC2-related kinase PITALRE is the catalytic subunit of active multimeric protein complexes. Biochem J. 1996 Oct 1;319(Pt 1):293–298. doi: 10.1042/bj3190293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Graña X., Reddy E. P. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 1995 Jul 20;11(2):211–219. [PubMed] [Google Scholar]
  12. Halevy O., Novitch B. G., Spicer D. B., Skapek S. X., Rhee J., Hannon G. J., Beach D., Lassar A. B. Correlation of terminal cell cycle arrest of skeletal muscle with induction of p21 by MyoD. Science. 1995 Feb 17;267(5200):1018–1021. doi: 10.1126/science.7863327. [DOI] [PubMed] [Google Scholar]
  13. Hateboer G., Kerkhoven R. M., Shvarts A., Bernards R., Beijersbergen R. L. Degradation of E2F by the ubiquitin-proteasome pathway: regulation by retinoblastoma family proteins and adenovirus transforming proteins. Genes Dev. 1996 Dec 1;10(23):2960–2970. doi: 10.1101/gad.10.23.2960. [DOI] [PubMed] [Google Scholar]
  14. Hattori M., Tugores A., Veloz L., Karin M., Brenner D. A. A simplified method for the preparation of transcriptionally active liver nuclear extracts. DNA Cell Biol. 1990 Dec;9(10):777–781. doi: 10.1089/dna.1990.9.777. [DOI] [PubMed] [Google Scholar]
  15. Ikeda M. A., Jakoi L., Nevins J. R. A unique role for the Rb protein in controlling E2F accumulation during cell growth and differentiation. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3215–3220. doi: 10.1073/pnas.93.8.3215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jiang H., Lin J., Young S. M., Goldstein N. I., Waxman S., Davila V., Chellappan S. P., Fisher P. B. Cell cycle gene expression and E2F transcription factor complexes in human melanoma cells induced to terminally differentiate. Oncogene. 1995 Sep 21;11(6):1179–1189. [PubMed] [Google Scholar]
  17. Johnson D. G. Regulation of E2F-1 gene expression by p130 (Rb2) and D-type cyclin kinase activity. Oncogene. 1995 Nov 2;11(9):1685–1692. [PubMed] [Google Scholar]
  18. Kiess M., Gill R. M., Hamel P. A. Expression and activity of the retinoblastoma protein (pRB)-family proteins, p107 and p130, during L6 myoblast differentiation. Cell Growth Differ. 1995 Oct;6(10):1287–1298. [PubMed] [Google Scholar]
  19. Lam E. W., Watson R. J. An E2F-binding site mediates cell-cycle regulated repression of mouse B-myb transcription. EMBO J. 1993 Jul;12(7):2705–2713. doi: 10.1002/j.1460-2075.1993.tb05932.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mayol X., Garriga J., Graña X. Cell cycle-dependent phosphorylation of the retinoblastoma-related protein p130. Oncogene. 1995 Aug 17;11(4):801–808. [PubMed] [Google Scholar]
  21. Mayol X., Graña X. pRB, p107 and p130 as transcriptional regulators: role in cell growth and differentiation. Prog Cell Cycle Res. 1997;3:157–169. doi: 10.1007/978-1-4615-5371-7_13. [DOI] [PubMed] [Google Scholar]
  22. Moberg K., Starz M. A., Lees J. A. E2F-4 switches from p130 to p107 and pRB in response to cell cycle reentry. Mol Cell Biol. 1996 Apr;16(4):1436–1449. doi: 10.1128/mcb.16.4.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohtani K., DeGregori J., Leone G., Herendeen D. R., Kelly T. J., Nevins J. R. Expression of the HsOrc1 gene, a human ORC1 homolog, is regulated by cell proliferation via the E2F transcription factor. Mol Cell Biol. 1996 Dec;16(12):6977–6984. doi: 10.1128/mcb.16.12.6977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Owens G. K. Regulation of differentiation of vascular smooth muscle cells. Physiol Rev. 1995 Jul;75(3):487–517. doi: 10.1152/physrev.1995.75.3.487. [DOI] [PubMed] [Google Scholar]
  25. Patel G., Kreider B., Rovera G., Reddy E. P. v-myb blocks granulocyte colony-stimulating factor-induced myeloid cell differentiation but not proliferation. Mol Cell Biol. 1993 Apr;13(4):2269–2276. doi: 10.1128/mcb.13.4.2269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Peterson C. A., Gordon H., Hall Z. W., Paterson B. M., Blau H. M. Negative control of the helix-loop-helix family of myogenic regulators in the NFB mutant. Cell. 1990 Aug 10;62(3):493–502. doi: 10.1016/0092-8674(90)90014-6. [DOI] [PubMed] [Google Scholar]
  27. Pickering J. G., Weir L., Rosenfield K., Stetz J., Jekanowski J., Isner J. M. Smooth muscle cell outgrowth from human atherosclerotic plaque: implications for the assessment of lesion biology. J Am Coll Cardiol. 1992 Nov 15;20(6):1430–1439. doi: 10.1016/0735-1097(92)90259-p. [DOI] [PubMed] [Google Scholar]
  28. Richon V. M., Lyle R. E., McGehee R. E., Jr Regulation and expression of retinoblastoma proteins p107 and p130 during 3T3-L1 adipocyte differentiation. J Biol Chem. 1997 Apr 11;272(15):10117–10124. doi: 10.1074/jbc.272.15.10117. [DOI] [PubMed] [Google Scholar]
  29. Schreiber E., Matthias P., Müller M. M., Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. doi: 10.1093/nar/17.15.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schwartz R. S. Neointima and arterial injury: dogs, rats, pigs, and more. Lab Invest. 1994 Dec;71(6):789–791. [PubMed] [Google Scholar]
  31. Schwartz S. M., deBlois D., O'Brien E. R. The intima. Soil for atherosclerosis and restenosis. Circ Res. 1995 Sep;77(3):445–465. doi: 10.1161/01.res.77.3.445. [DOI] [PubMed] [Google Scholar]
  32. Shin E. K., Shin A., Paulding C., Schaffhausen B., Yee A. S. Multiple change in E2F function and regulation occur upon muscle differentiation. Mol Cell Biol. 1995 Apr;15(4):2252–2262. doi: 10.1128/mcb.15.4.2252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith E. J., Leone G., DeGregori J., Jakoi L., Nevins J. R. The accumulation of an E2F-p130 transcriptional repressor distinguishes a G0 cell state from a G1 cell state. Mol Cell Biol. 1996 Dec;16(12):6965–6976. doi: 10.1128/mcb.16.12.6965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Steer C. J. Liver regeneration. FASEB J. 1995 Nov;9(14):1396–1400. doi: 10.1096/fasebj.9.14.7589980. [DOI] [PubMed] [Google Scholar]
  35. Tommasi S., Pfeifer G. P. In vivo structure of the human cdc2 promoter: release of a p130-E2F-4 complex from sequences immediately upstream of the transcription initiation site coincides with induction of cdc2 expression. Mol Cell Biol. 1995 Dec;15(12):6901–6913. doi: 10.1128/mcb.15.12.6901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vairo G., Livingston D. M., Ginsberg D. Functional interaction between E2F-4 and p130: evidence for distinct mechanisms underlying growth suppression by different retinoblastoma protein family members. Genes Dev. 1995 Apr 1;9(7):869–881. doi: 10.1101/gad.9.7.869. [DOI] [PubMed] [Google Scholar]
  37. Valtieri M., Tweardy D. J., Caracciolo D., Johnson K., Mavilio F., Altmann S., Santoli D., Rovera G. Cytokine-dependent granulocytic differentiation. Regulation of proliferative and differentiative responses in a murine progenitor cell line. J Immunol. 1987 Jun 1;138(11):3829–3835. [PubMed] [Google Scholar]
  38. Wei G. L., Krasinski K., Kearney M., Isner J. M., Walsh K., Andrés V. Temporally and spatially coordinated expression of cell cycle regulatory factors after angioplasty. Circ Res. 1997 Mar;80(3):418–426. [PubMed] [Google Scholar]
  39. Williams C. D., Linch D. C., Sørensen T. S., La Thangue N. B., Thomas N. S. The predominant E2F complex in human primary haemopoietic cells and in AML blasts contains E2F-4, DP-1 and p130. Br J Haematol. 1997 Mar;96(4):688–696. doi: 10.1046/j.1365-2141.1997.d01-2086.x. [DOI] [PubMed] [Google Scholar]
  40. Wolf D. A., Hermeking H., Albert T., Herzinger T., Kind P., Eick D. A complex between E2F and the pRb-related protein p130 is specifically targeted by the simian virus 40 large T antigen during cell transformation. Oncogene. 1995 Jun 1;10(11):2067–2078. [PubMed] [Google Scholar]
  41. Zhu L., Zhu L., Xie E., Chang L. S. Differential roles of two tandem E2F sites in repression of the human p107 promoter by retinoblastoma and p107 proteins. Mol Cell Biol. 1995 Jul;15(7):3552–3562. doi: 10.1128/mcb.15.7.3552. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES