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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Sep 13;91(19):9101–9105. doi: 10.1073/pnas.91.19.9101

Enhancer 1 binding factor (E1BF), a Ku-related protein, is a growth-regulated RNA polymerase I transcription factor: association of a repressor activity with purified E1BF from serum-deprived cells.

H Niu 1, S T Jacob 1
PMCID: PMC44755  PMID: 8090777

Abstract

Previous studies from this laboratory have demonstrated that the enhancer 1 binding factor (E1BF), a Ku-related protein, purified from the serum-enriched cells functions as a positive factor in an RNA polymerase (pol I) transcription system. We have now shown that E1BF purified from the serum-deprived cells (E1BFs) can inhibit rDNA transcription completely in a fractionated extract from the cells grown in serum-enriched medium. The suppression of transcription was overcome by the addition of control E1BF (E1BFc). Immunoprecipitation of purified E1BFs by the anti-Ku monoclonal antibody and addition of the supernatant to the transcription reaction mixture prevented the inhibition significantly, whereas immunoprecipitation with the control mouse IgG did not restore the transcription. The transcriptional repressor activity associated with the final DNA affinity column fractions copurified with E1BF. Neither the amount of E1BF nor its promoter binding activity was altered following serum depletion. E1BFs selectively inhibited the initiation of rDNA transcription. The inhibitory activity of E1BFs was not due to a nonspecific RNase activity. These data suggest that E1BF is post-translationally modified following serum starvation of cells, and that the repressor activity of E1BFs is largely responsible for the down-regulation of pol I transcription in serum-deprived cells.

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Selected References

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  1. Bateman E., Paule M. R. Regulation of eukaryotic ribosomal RNA transcription by RNA polymerase modification. Cell. 1986 Nov 7;47(3):445–450. doi: 10.1016/0092-8674(86)90601-x. [DOI] [PubMed] [Google Scholar]
  2. Cavanaugh A. H., Gokal P. K., Lawther R. P., Thompson E. A., Jr Glucocorticoid inhibition of initiation of transcription of the DNA encoding rRNA (rDNA) in lymphosarcoma P1798 cells. Proc Natl Acad Sci U S A. 1984 Feb;81(3):718–721. doi: 10.1073/pnas.81.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Garg L. C., Dixit A., Jacob S. T. A 37-base pair element in the far upstream spacer region can enhance transcription of rat rDNA in vitro and can bind to the core promoter-binding factor(s). J Biol Chem. 1989 Jan 5;264(1):220–224. [PubMed] [Google Scholar]
  4. Ghosh A. K., Hoff C. M., Jacob S. T. Characterization of the 130-bp repeat enhancer element of the rat ribosomal gene: functional interaction with transcription factor E1BF. Gene. 1993 Mar 30;125(2):217–222. doi: 10.1016/0378-1119(93)90332-w. [DOI] [PubMed] [Google Scholar]
  5. Gottesfeld J. M., Wolf V. J., Dang T., Forbes D. J., Hartl P. Mitotic repression of RNA polymerase III transcription in vitro mediated by phosphorylation of a TFIIIB component. Science. 1994 Jan 7;263(5143):81–84. doi: 10.1126/science.8272869. [DOI] [PubMed] [Google Scholar]
  6. Hoff C. M., Ghosh A. K., Prabhakar B. S., Jacob S. T. Enhancer 1 binding factor, a Ku-related protein, is a positive regulator of RNA polymerase I transcription initiation. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):762–766. doi: 10.1073/pnas.91.2.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hoff C. M., Jacob S. T. Characterization of the factor E1BF from a rat hepatoma that modulates ribosomal RNA gene transcription and its relationship to the human Ku autoantigen. Biochem Biophys Res Commun. 1993 Feb 15;190(3):747–753. doi: 10.1006/bbrc.1993.1112. [DOI] [PubMed] [Google Scholar]
  8. Jacob S. T., Sajdel E. M., Munro H. N. Regulation of nucleolar RNA metabolism by hydrocortisone. Eur J Biochem. 1969 Feb;7(4):449–453. doi: 10.1111/j.1432-1033.1969.tb19630.x. [DOI] [PubMed] [Google Scholar]
  9. Kermekchiev M., Muramatsu M. Presence of an inhibitor of RNA polymerase I mediated transcription in extracts from growth arrested mouse cells. Nucleic Acids Res. 1993 Feb 11;21(3):447–453. doi: 10.1093/nar/21.3.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kuhn A., Stefanovsky V., Grummt I. The nucleolar transcription activator UBF relieves Ku antigen-mediated repression of mouse ribosomal gene transcription. Nucleic Acids Res. 1993 May 11;21(9):2057–2063. doi: 10.1093/nar/21.9.2057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Liu X., Miller C. W., Koeffler P. H., Berk A. J. The p53 activation domain binds the TATA box-binding polypeptide in Holo-TFIID, and a neighboring p53 domain inhibits transcription. Mol Cell Biol. 1993 Jun;13(6):3291–3300. doi: 10.1128/mcb.13.6.3291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mahajan P. B., Thompson E. A. Hormonal regulation of transcription of rDNA. Purification and characterization of the hormone-regulated transcription factor IC. J Biol Chem. 1990 Sep 25;265(27):16225–16233. [PubMed] [Google Scholar]
  13. Muramatsu M., Shimada N., Higashinakagawa T. Effect of cycloheximide on the nucleolar RNA synthesis in rat liver. J Mol Biol. 1970 Oct 14;53(1):91–106. doi: 10.1016/0022-2836(70)90047-1. [DOI] [PubMed] [Google Scholar]
  14. O'Mahony D. J., Xie W. Q., Smith S. D., Singer H. A., Rothblum L. I. Differential phosphorylation and localization of the transcription factor UBF in vivo in response to serum deprivation. In vitro dephosphorylation of UBF reduces its transactivation properties. J Biol Chem. 1992 Jan 5;267(1):35–38. [PubMed] [Google Scholar]
  15. Paule M. R. Polymerase I transcription, termination, and processing. Gene Expr. 1993;3(1):1–9. [PMC free article] [PubMed] [Google Scholar]
  16. Sauer F., Jäckle H. Dimerization and the control of transcription by Krüppel. Nature. 1993 Jul 29;364(6436):454–457. doi: 10.1038/364454a0. [DOI] [PubMed] [Google Scholar]
  17. Schnapp A., Pfleiderer C., Rosenbauer H., Grummt I. A growth-dependent transcription initiation factor (TIF-IA) interacting with RNA polymerase I regulates mouse ribosomal RNA synthesis. EMBO J. 1990 Sep;9(9):2857–2863. doi: 10.1002/j.1460-2075.1990.tb07475.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schnapp A., Schnapp G., Erny B., Grummt I. Function of the growth-regulated transcription initiation factor TIF-IA in initiation complex formation at the murine ribosomal gene promoter. Mol Cell Biol. 1993 Nov;13(11):6723–6732. doi: 10.1128/mcb.13.11.6723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tower J., Sollner-Webb B. Transcription of mouse rDNA is regulated by an activated subform of RNA polymerase I. Cell. 1987 Sep 11;50(6):873–883. doi: 10.1016/0092-8674(87)90514-9. [DOI] [PubMed] [Google Scholar]
  20. Vallett S. M., Brudnak M., Pellegrini M., Weber H. W. In vivo regulation of rRNA transcription occurs rapidly in nondividing and dividing Drosophila cells in response to a phorbol ester and serum. Mol Cell Biol. 1993 Feb;13(2):928–933. doi: 10.1128/mcb.13.2.928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Voit R., Schnapp A., Kuhn A., Rosenbauer H., Hirschmann P., Stunnenberg H. G., Grummt I. The nucleolar transcription factor mUBF is phosphorylated by casein kinase II in the C-terminal hyperacidic tail which is essential for transactivation. EMBO J. 1992 Jun;11(6):2211–2218. doi: 10.1002/j.1460-2075.1992.tb05280.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Webb M. L., Mealey-Cavender J. F., Jacob S. T. Glucocorticoid-induced stimulation of ribosomal gene transcription in rat hepatoma cells is mediated by modification of RNA polymerase I or an associated factor. Mol Endocrinol. 1989 Nov;3(11):1861–1868. doi: 10.1210/mend-3-11-1861. [DOI] [PubMed] [Google Scholar]
  23. Yaneva M., Jhiang S. Expression of the Ku protein during cell proliferation. Biochim Biophys Acta. 1991 Oct 8;1090(2):181–187. doi: 10.1016/0167-4781(91)90099-8. [DOI] [PubMed] [Google Scholar]
  24. Zhang J., Jacob S. T. Purification and characterization of a novel factor which stimulates rat ribosomal gene transcription in vitro by interacting with enhancer and core promoter elements. Mol Cell Biol. 1990 Oct;10(10):5177–5186. doi: 10.1128/mcb.10.10.5177. [DOI] [PMC free article] [PubMed] [Google Scholar]

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