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. 1992 Mar 25;20(6):1243–1249. doi: 10.1093/nar/20.6.1243

The rat poly pyrimidine tract binding protein (PTB) interacts with a single-stranded DNA motif in a liver-specific enhancer.

P Jansen-Dürr 1, M Boshart 1, B Lupp 1, A Bosserhoff 1, R W Frank 1, G Schütz 1
PMCID: PMC312165  PMID: 1561080

Abstract

We characterized and purified a protein from rat liver which specifically binds to a DNA motif present in a liver-specific enhancer of the rat tyrosine aminotransferase (TAT) gene, when offered as single-stranded DNA. Binding is highly sequence-specific and coincides with a region known to be essential for function of the enhancer. Microsequencing revealed that this protein is the rat homologue of the mouse and human poly Pyrimidine Tract binding protein (PTB), which has been shown to bind to premRNA and may participate in RNA splicing. This finding was corroborated by subsequent Western blot experiments using a PTB-specific antibody. These findings indicate a possible dual role for this protein in RNA processing and transcription.

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  1. Baeuerle P. A., Baltimore D. A 65-kappaD subunit of active NF-kappaB is required for inhibition of NF-kappaB by I kappaB. Genes Dev. 1989 Nov;3(11):1689–1698. doi: 10.1101/gad.3.11.1689. [DOI] [PubMed] [Google Scholar]
  2. Boeuf H., Reimund B., Jansen-Durr P., Kédinger C. Differential activation of the E2F transcription factor by the adenovirus EIa and EIV products in F9 cells. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1782–1786. doi: 10.1073/pnas.87.5.1782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boshart M., Weih F., Schmidt A., Fournier R. E., Schütz G. A cyclic AMP response element mediates repression of tyrosine aminotransferase gene transcription by the tissue-specific extinguisher locus Tse-1. Cell. 1990 Jun 1;61(5):905–916. doi: 10.1016/0092-8674(90)90201-o. [DOI] [PubMed] [Google Scholar]
  4. Bosserhoff A., Wallach J., Frank R. W. Micropreparative separation of peptides derived from sodium dodecyl sulphate-solubilized proteins. J Chromatogr. 1989 Jun 28;473(1):71–77. doi: 10.1016/s0021-9673(00)91291-3. [DOI] [PubMed] [Google Scholar]
  5. Brunel F., Alzari P. M., Ferrara P., Zakin M. M. Cloning and sequencing of PYBP, a pyrimidine-rich specific single strand DNA-binding protein. Nucleic Acids Res. 1991 Oct 11;19(19):5237–5245. doi: 10.1093/nar/19.19.5237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brunel F., Ochoa A., Schaeffer E., Boissier F., Guillou Y., Cereghini S., Cohen G. N., Zakin M. M. Interactions of DNA-binding proteins with the 5' region of the human transferrin gene. J Biol Chem. 1988 Jul 25;263(21):10180–10185. [PubMed] [Google Scholar]
  7. Brunelle A., Schleif R. F. Missing contact probing of DNA-protein interactions. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6673–6676. doi: 10.1073/pnas.84.19.6673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cox M. M., Lehman I. R. Enzymes of general recombination. Annu Rev Biochem. 1987;56:229–262. doi: 10.1146/annurev.bi.56.070187.001305. [DOI] [PubMed] [Google Scholar]
  9. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dynan W. S. Modularity in promoters and enhancers. Cell. 1989 Jul 14;58(1):1–4. doi: 10.1016/0092-8674(89)90393-0. [DOI] [PubMed] [Google Scholar]
  11. Gaillard C., Strauss F. Sequence-specific single-strand-binding protein for the simian virus 40 early promoter stimulates transcription in vitro. J Mol Biol. 1990 Sep 20;215(2):245–255. doi: 10.1016/S0022-2836(05)80343-2. [DOI] [PubMed] [Google Scholar]
  12. García-Blanco M. A., Jamison S. F., Sharp P. A. Identification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns. Genes Dev. 1989 Dec;3(12A):1874–1886. doi: 10.1101/gad.3.12a.1874. [DOI] [PubMed] [Google Scholar]
  13. Gil A., Sharp P. A., Jamison S. F., Garcia-Blanco M. A. Characterization of cDNAs encoding the polypyrimidine tract-binding protein. Genes Dev. 1991 Jul;5(7):1224–1236. doi: 10.1101/gad.5.7.1224. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Johnson P. F., McKnight S. L. Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989;58:799–839. doi: 10.1146/annurev.bi.58.070189.004055. [DOI] [PubMed] [Google Scholar]
  16. Lannigan D. A., Notides A. C. Estrogen receptor selectively binds the "coding strand" of an estrogen responsive element. Proc Natl Acad Sci U S A. 1989 Feb;86(3):863–867. doi: 10.1073/pnas.86.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  18. Mukherjee R., Chambon P. A single-stranded DNA-binding protein promotes the binding of the purified oestrogen receptor to its responsive element. Nucleic Acids Res. 1990 Oct 11;18(19):5713–5716. doi: 10.1093/nar/18.19.5713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nitsch D., Stewart A. F., Boshart M., Mestril R., Weih F., Schütz G. Chromatin structures of the rat tyrosine aminotransferase gene relate to the function of its cis-acting elements. Mol Cell Biol. 1990 Jul;10(7):3334–3342. doi: 10.1128/mcb.10.7.3334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Patton J. G., Mayer S. A., Tempst P., Nadal-Ginard B. Characterization and molecular cloning of polypyrimidine tract-binding protein: a component of a complex necessary for pre-mRNA splicing. Genes Dev. 1991 Jul;5(7):1237–1251. doi: 10.1101/gad.5.7.1237. [DOI] [PubMed] [Google Scholar]
  21. Santoro C., Mermod N., Andrews P. C., Tjian R. A family of human CCAAT-box-binding proteins active in transcription and DNA replication: cloning and expression of multiple cDNAs. Nature. 1988 Jul 21;334(6179):218–224. doi: 10.1038/334218a0. [DOI] [PubMed] [Google Scholar]
  22. Santoro I. M., Yi T. M., Walsh K. Identification of single-stranded-DNA-binding proteins that interact with muscle gene elements. Mol Cell Biol. 1991 Apr;11(4):1944–1953. doi: 10.1128/mcb.11.4.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wang J., Pederson T. A 62,000 molecular weight spliceosome protein crosslinks to the intron polypyrimidine tract. Nucleic Acids Res. 1990 Oct 25;18(20):5995–6001. doi: 10.1093/nar/18.20.5995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wilkison W. O., Min H. Y., Claffey K. P., Satterberg B. L., Spiegelman B. M. Control of the adipsin gene in adipocyte differentiation. Identification of distinct nuclear factors binding to single- and double-stranded DNA. J Biol Chem. 1990 Jan 5;265(1):477–482. [PubMed] [Google Scholar]

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