Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

RNA polymerase I transcription in confluent cells: Rb downregulates rDNA transcription during confluence-induced cell cycle arrest

Oncogene volume 19, pages 3487–3497 (2000)Cite this article

Abstract

When 3T6 cells are confluent, they withdraw from the cell cycle. Concomitant with cell cycle arrest a significant reduction in RNA polymerase I transcription (80% decrease at 100% confluence) is observed. In the present study, we examined mechanism(s) through which transcription of the ribosomal genes is coupled to cell cycle arrest induced by cell density. Interestingly with an increase in cell density (from 3–43% confluence), a significant accumulation in the cellular content of hyperphosphorylated Rb was observed. As cell density increased further, the hypophosphorylated form of Rb became predominant and accumulated in the nucleoli. Co-immunoprecipitation experiments demonstrated there was also a significant rise in the amount of hypophosphorylated Rb associated with the rDNA transcription factor UBF. This increased interaction between Rb and UBF correlated with the reduced rate of rDNA transcription. Furthermore, overexpression of recombinant Rb inhibited UBF-dependent activation of transcription from a cotransfected rDNA reporter in either confluent or exponential cells. The amounts or activities of the rDNA transcription components we examined did not significantly change with cell cycle arrest. Although the content of PAF53, a polymerase associated factor, was altered marginally (decreased 38%), the time course and magnitude of the decrease did not correlate with the reduced rate of rDNA transcription. The results presented support a model wherein regulation of the binding of UBF to Rb and, perhaps the cellular content of PAF53, are components of the mechanism through which cell cycle and rDNA transcription are linked.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

Abbreviations

PAF53:

polymerase associated factor 53

PBS:

phosphate-buffered saline

Rb:

retinoblastoma susceptibility gene product

rDNA:

ribosomal DNA

rRNA:

ribosomal RNA

SL-1:

selectivity factor 1

TFIC:

transcription factor 1C

UBF:

upstream binding factor

References

  • Alberts B, Bray D, Lewis J, Raff M, Roberts K and Watson JD. (eds). . 1994 Molecular Biology of the Cell. Garland Publishing Inc.: New York, USA.

    Google Scholar 

  • Ausubel F, Brent R, Kingston R, Moore D, Seidman J, Smith J and Struhl K. (eds). . 1993 Current Protocols in Molecular Biology. Wiley Interscience: USA.

    Google Scholar 

  • Beckmann H, Chen JL, O'Brien T and Tjian R. . 1995 Science 270: 1506–1509.

  • Brun RP, Ryan K and Sollner-Webb B. . 1994 Mol. Cell. Biol. 14: 5010–5021.

  • Cavanaugh AH, Gokal PK, Lawther RP and Thompson EA. . 1984 Proc. Natl. Acad. Sci. USA 81: 718–721.

  • Cavanaugh AH, Hempel WM, Taylor LJ, Rogalsky V, Todorov G and Rothblum LI. . 1995 Nature 374: 177–180.

  • Cesarone CF, Bolognesi C and Santi L. . 1979 Anal. Biochem. 100: 188–197.

  • Cobrinik D, Dowdy SF, Hinds PW, Mittnacht S and Weinberg RA. . 1992 Trends Biochem. Science 17: 312–315.

  • Doree M and Galas S. . 1994 FASEB J. 8: 1114–1121.

  • Glibetic M, Larson D, Taylor L, Hannan R, Sells B and Rothblum L. . 1995 J. Biol. Chem. 270: 4209–4212.

  • Goodrich DW, Wang NP, Qian Y-W, Lee EY-H and Lee W-H.. 1991 Cell 67: 293–302.

  • Haglund RE and Rothblum LI. . 1987 Mol. Cell. Biochem. 73: 11–25.

  • Hamel PA, Gallie BL and Phillips RA. . 1992 Trends Genetics 8: 180–185.

  • Hanada K, Song CZ, Tamamoto K, Yano K, Maeda Y, Yamaguchi K and Muramatsu M. . 1996 EMBO J. 15: 2217–2226.

  • Hannan KM, Hannan RD and Rothblum LI. . 1998a Frontiers Biosci. 3: 376–398.

  • Hannan KM, Hannan RD, Smith SD, Hempel WM, Jefferson LS and Rothblum LI. . 1999a Oncogene submitted.

  • Hannan RD, Stennard FA and West AK. . 1993 Mol. Cell. Cardiol. 25: 1137–1148.

  • Hannan RD and Rothblum LI. . 1995 Cardiovas. Res. 30: 501–510.

  • Hannan RD, Luyken J and Rothblum LI. . 1995 J. Biol. Chem. 270: 8290–8297.

  • Hannan RD, Stefanovsky V, Taylor L, Moss T and Rothblum LI. . 1996b Proc. Natl. Acad. Sci. USA 93: 8750–8755.

  • Hannan RD, Luyken J and Rothblum LI. . 1996a J. Biol. Chem. 271: 3213–3220.

  • Hannan RD, Hempel W, Cavanaugh A, Arino T, Dimitrov SI, Moss T and Rothblum LI. . 1998b J. Biol. Chem. 273: 1257–1267.

  • Hannan RD, Stefanovsky V, Arino T, Rothblum LI and Moss T. . 1999b Nucleic Acids. Res. 27: 1205–1213.

  • Heix J, Vente A, Voit R, Budde A, Michaelidis M and Grummt I. . 1998 EMBO J. 17: 7373–7381.

  • Hempel WM, Cavanaugh A, Hannan R, Taylor L and Rothblum LI. . 1996 Mol. Cell. Biol. 16: 557–563.

  • Kihm AJ, Hershey JC, Haystead TAJ, Madsen CS and Owens GK. . 1998 Proc. Natl. Acad. Sci. 95: 14816–14820.

  • Kuhn A, Voit R, Stefanovsky R, Evers R, Bianchi M and Grummt I. . 1994 EMBO J. 13: 416–424.

  • Larson DE, Xie W, Glibetic M, O'Mahony D, Sells B and Rothblum LI. . 1993 Proc. Natl. Acad. Sci. USA 90: 7933–7936.

  • Learned RS, Cordes S and Tjian R. . 1985 Mol. Cell. Biol. 5: 1358–1369.

  • Luyken J, Hannan RD, Cheung J and Rothblum LI. . 1995 Circ. Res. 78: 354–361.

  • Mauck JC and Green H. . 1973 Proc. Natl. Acad. Sci. USA 70: 2819–2822.

  • Mahajan PB and Thompson EA. . 1990 J. Biol. Chem. 265: 16225–16233.

  • Miesfeld R and Arnheim N. . 1984 Mol. Cell. Biol. 4: 221–227.

  • Mihara K, Cao X, Yen A, Chandler S, Driscoll B, Murphree A, T'ang A and Fung YK. . 1989 Science 246: 1300–1303.

  • Mishima Y, Financsek I, Kominami R and Muramatsu M. . 1982 Nucleic Acids Res. 10: 6659–6670.

  • Moss T and Stefanovsky VY. . 1995 Prog. Nucl. Acids Res. Mol. Biol. 50: 25–66.

  • Nevins JR. . 1992 Science 258: 424–429.

  • O'Mahony DJ and Rothblum LI. . 1991 Proc. Natl. Acad. Sci. USA 88: 3180–3184.

  • O'Mahony DJ, Xie W, Smith S, Singer H and Rothblum LI. . 1992 J. Biol. Chem. 267: 35–38.

  • Palmer TD, Miller A, Reeder RH and McStay B. . 1993 Nucleic Acids Res. 21: 3451–3457.

  • Pardee AB. . 1989 Science 246: 603–608.

  • Paule M. (ed). . 1994 Transcription: Mechanisms and Regulation. Raven Press Ltd.: NY.

    Google Scholar 

  • Robbins PD, Horowitz JM and Mulligan RC. . 1990 Nature 346: 668–671.

  • Schnapp A, Pfleiderer C, Rosenbauer H and Grummt I. . 1990 EMBO J. 9: 2857–2863.

  • Schnapp A, Schnapp G, Erny B and Grummt I. . 1993 Mol. Cell. Biol. 13: 6723–6732.

  • Schnapp G, Santori F, Carles C, Riva M and Grummt I. . 1994 EMBO J. 13: 190–199.

  • Sherr CJ. . 1993 Cell 73: 1059–1065.

  • Smith SD, Oriahi E, Lowe D, Yang-Yen HF, O'Mahony D, Rose K, Chen C and Rothblum LI. . 1990 Mol. Cell. Biol. 10: 3105–3116.

  • Smith SD, O'Mahony D, Kinsella BT and Rothblum LI. . 1993 Gene Expression 3: 229–236.

  • Voit R, Schnapp A, Kuhn A, Rosenbauer H, Hirschmann P, Stunnenberg HG and Grummt I. . 1992 EMBO J. 11: 2211–2218.

  • Voit R, Schafer K and Grummt I. . 1997 Mol. Cell. Biol. 17: 4230–4237.

  • Voit R, Hoffmann M and Grummt I. . 1999 EMBO J. 18: 1891–1899.

Download references

Acknowledgements

This study was supported in part by National Institute of Health grants GM48991 (LI Rothblum), DK15658 (LS Jefferson) and DK13499 (LS Jefferson), the Juvenile Diabetes Foundation JDFI195051 (LS Jefferson) and an award from the Geisinger Foundation (LI Rothblum). All imaging experiments were performed at the Whitehead Institute Microscopy Facility. BK Kennedy is supported by a Leukemia Society of America Postdoctoral Fellowship. Dr Muramatsu of the Saitama Medical School of Japan, Department of Biochemistry graciously provided the original sample of PAF53 antisera used in these studies.

Author information

Author notes

  1. Katherine M Hannan & Ross D Hannan

    Present address: Baker Medical Research Institute, P.O. Box 6492, Melbourne, 8008, Victoria, Australia

Authors and Affiliations

  1. Department of Cellular and Molecular Physiology, The Pennsylvania State University, College of Medicine, Hershey, 17033, Pennsylvania, PA, USA

    Katherine M Hannan & Leonard S Jefferson

  2. Department of Molecular Oncology, Massachusetts General Hospital Cancer Center, Building 149, 13th Street, Charlestown, 02129, Massachusetts, MA, USA

    Brian K Kennedy

  3. The Henry Hood Research Program, Sigfried and Janet Weis Center for Research, The Pennsylvania State University College of Medicine, Danville, 17822-2618, Pennsylvania, PA, USA

    Katherine M Hannan, Alice H Cavanaugh, Ross D Hannan, Iwona Hirschler-Laszkiewicz & Lawrence I Rothblum

Authors
  1. Katherine M Hannan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brian K Kennedy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alice H Cavanaugh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ross D Hannan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Iwona Hirschler-Laszkiewicz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Leonard S Jefferson
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lawrence I Rothblum
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hannan, K., Kennedy, B., Cavanaugh, A. et al. RNA polymerase I transcription in confluent cells: Rb downregulates rDNA transcription during confluence-induced cell cycle arrest. Oncogene 19, 3487–3497 (2000). https://doi.org/10.1038/sj.onc.1203690

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1203690

Keywords

This article is cited by

Search

Advanced search

Quick links