Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1989 Aug 25;17(16):6693–6711. doi: 10.1093/nar/17.16.6693

Multiple protein binding sites within the ovalbumin gene 5'-flanking region: isolation and characterization of sequence-specific binding proteins.

M Pastorcic 1, M K Bagchi 1, S Y Tsai 1, M J Tsai 1, B W O'Malley 1
PMCID: PMC318360  PMID: 2780293

Abstract

To understand the molecular basis of the steroid hormone regulated expression of the ovalbumin gene, we sought to identify and isolate nuclear factors from chicken oviduct which interact specifically with the ovalbumin promoter. Using DNase I footprinting and mobility shift assays, we have defined at least four distinct protein binding sites, OV-150, OV-220, OV-250 and OV-330, in the promoter region between -100 to -400. Binding competition and protein fractionation studies revealed the existence of two distinct proteins, each recognizing two promoter sites: Both OV-330 and OV-250 are recognized by one protein factor which is distinct from the one binding to both OV-220 and OV-150. The location of the DNase I footprints coincides with those of in vivo chromatin hypersensitive sites. The OV-330 site is located in a sequence area required for the repression of the gene in the absence of hormone. The factor binding to OV-330 has been substantially purified and renaturation experiments indicate that the binding activity is associated with a polypeptide(s) of Mr 40K.

Full text

PDF
6693

Images in this article

6696Image
on p.6696
6697Image
on p.6697
6699Image
on p.6699
6700Image
on p.6700
6704Image
on p.6704
6706Image
on p.6706

Selected References

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

  1. Bagchi M. K., Tsai S. Y., Tsai M. J., O'Malley B. W. Purification and characterization of chicken ovalbumin gene upstream promoter transcription factor from homologous oviduct cells. Mol Cell Biol. 1987 Dec;7(12):4151–4158. doi: 10.1128/mcb.7.12.4151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradshaw M. S., Tsai M. J., O'Malley B. W. A far upstream ovalbumin enhancer binds nuclear factor-1-like factor. J Biol Chem. 1988 Jun 15;263(17):8485–8490. [PubMed] [Google Scholar]
  3. Briggs M. R., Kadonaga J. T., Bell S. P., Tjian R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science. 1986 Oct 3;234(4772):47–52. doi: 10.1126/science.3529394. [DOI] [PubMed] [Google Scholar]
  4. Corthésy B., Hipskind R., Theulaz I., Wahli W. Estrogen-dependent in vitro transcription from the vitellogenin promoter in liver nuclear extracts. Science. 1988 Mar 4;239(4844):1137–1139. doi: 10.1126/science.2830672. [DOI] [PubMed] [Google Scholar]
  5. Dierich A., Gaub M. P., LePennec J. P., Astinotti D., Chambon P. Cell-specificity of the chicken ovalbumin and conalbumin promoters. EMBO J. 1987 Aug;6(8):2305–2312. doi: 10.1002/j.1460-2075.1987.tb02505.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Garner M. M., Revzin A. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981 Jul 10;9(13):3047–3060. doi: 10.1093/nar/9.13.3047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gaub M. P., Dierich A., Astinotti D., Touitou I., Chambon P. The chicken ovalbumin promoter is under negative control which is relieved by steroid hormones. EMBO J. 1987 Aug;6(8):2313–2320. doi: 10.1002/j.1460-2075.1987.tb02506.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gerlinger P., Krust A., LeMeur M., Perrin F., Cochet M., Gannon F., Dupret D., Chambon P. Multiple initiation and polyadenylation sites for the chicken ovomucoid transcription unit. J Mol Biol. 1982 Dec 5;162(2):345–364. doi: 10.1016/0022-2836(82)90531-9. [DOI] [PubMed] [Google Scholar]
  9. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  10. Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
  11. Hager L. J., McKnight G. S., Palmiter R. D. Glucocorticoid induction of egg white mRNAs in chick oviduct. J Biol Chem. 1980 Aug 25;255(16):7796–7800. [PubMed] [Google Scholar]
  12. Horikoshi M., Carey M. F., Kakidani H., Roeder R. G. Mechanism of action of a yeast activator: direct effect of GAL4 derivatives on mammalian TFIID-promoter interactions. Cell. 1988 Aug 26;54(5):665–669. doi: 10.1016/s0092-8674(88)80011-4. [DOI] [PubMed] [Google Scholar]
  13. Jantzen H. M., Strähle U., Gloss B., Stewart F., Schmid W., Boshart M., Miksicek R., Schütz G. Cooperativity of glucocorticoid response elements located far upstream of the tyrosine aminotransferase gene. Cell. 1987 Apr 10;49(1):29–38. doi: 10.1016/0092-8674(87)90752-5. [DOI] [PubMed] [Google Scholar]
  14. Kaye J. S., Pratt-Kaye S., Bellard M., Dretzen G., Bellard F., Chambon P. Steroid hormone dependence of four DNase I-hypersensitive regions located within the 7000-bp 5'-flanking segment of the ovalbumin gene. EMBO J. 1986 Feb;5(2):277–285. doi: 10.1002/j.1460-2075.1986.tb04210.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Knoll B. J., Zarucki-Schulz T., Dean D. C., O'Malley B. W. Definition of the ovalbumin gene promoter by transfer of an ovalglobin fusion gene into cultured cells. Nucleic Acids Res. 1983 Oct 11;11(19):6733–6754. doi: 10.1093/nar/11.19.6733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  17. O'Malley B. W., Roop D. R., Lai E. C., Nordstrom J. L., Catterall J. F., Swaneck G. E., Colbert D. A., Tsai M. J., Dugaiczyk A., Woo S. L. The ovalbumin gene: organization, structure, transcription, and regulation. Recent Prog Horm Res. 1979;35:1–46. doi: 10.1016/b978-0-12-571135-7.50005-9. [DOI] [PubMed] [Google Scholar]
  18. Pastorcic M., Wang H., Elbrecht A., Tsai S. Y., Tsai M. J., O'Malley B. W. Control of transcription initiation in vitro requires binding of a transcription factor to the distal promoter of the ovalbumin gene. Mol Cell Biol. 1986 Aug;6(8):2784–2791. doi: 10.1128/mcb.6.8.2784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sagami I., Tsai S. Y., Wang H., Tsai M. J., O'Malley B. W. Identification of two factors required for transcription of the ovalbumin gene. Mol Cell Biol. 1986 Dec;6(12):4259–4267. doi: 10.1128/mcb.6.12.4259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sawadogo M., Roeder R. G. Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell. 1985 Nov;43(1):165–175. doi: 10.1016/0092-8674(85)90021-2. [DOI] [PubMed] [Google Scholar]
  21. Slater E. P., Rabenau O., Karin M., Baxter J. D., Beato M. Glucocorticoid receptor binding and activation of a heterologous promoter by dexamethasone by the first intron of the human growth hormone gene. Mol Cell Biol. 1985 Nov;5(11):2984–2992. doi: 10.1128/mcb.5.11.2984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tora L., Gronemeyer H., Turcotte B., Gaub M. P., Chambon P. The N-terminal region of the chicken progesterone receptor specifies target gene activation. Nature. 1988 May 12;333(6169):185–188. doi: 10.1038/333185a0. [DOI] [PubMed] [Google Scholar]
  23. Tsai S. Y., Sagami I., Wang H., Tsai M. J., O'Malley B. W. Interactions between a DNA-binding transcription factor (COUP) and a non-DNA binding factor (S300-II). Cell. 1987 Aug 28;50(5):701–709. doi: 10.1016/0092-8674(87)90328-x. [DOI] [PubMed] [Google Scholar]
  24. Tsai S. Y., Tsai M. J., O'Malley B. W. Specific 5' flanking sequences are required for faithful initiation of in vitro transcription of the ovalbumin gene. Proc Natl Acad Sci U S A. 1981 Feb;78(2):879–883. doi: 10.1073/pnas.78.2.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang L. H., Tsai S. Y., Sagami I., Tsai M. J., O'Malley B. W. Purification and characterization of chicken ovalbumin upstream promoter transcription factor from HeLa cells. J Biol Chem. 1987 Nov 25;262(33):16080–16086. [PubMed] [Google Scholar]
  26. Zarucki-Schulz T., Tsai S. Y., Itakura K., Soberon X., Wallace R. B., Tsai M. J., Woo S. L., O'Malley B. W. Point mutagenesis of the ovalbumin gene promoter sequence and its effect on in vitro transcription. J Biol Chem. 1982 Sep 25;257(18):11070–11077. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES