Abstract
A new methodology for the identification of genes modulated by transcription factors in vivo is described. Mouse genomic DNA fragments bound by the thyroid hormone receptor (T3R) were selected and amplified in vitro. Subsequent hybridisation with biotinylated cDNA allowed the selection of those DNA fragments containing binding sites for T3R that corresponded to transcribed DNA. Expression analysis of the corresponding genes showed that more than 80% are indeed modulated by thyroid hormones in vivo in the liver. Together with the presence of consensus binding sites for T3R this result suggests that the selected DNA fragments may contain T3R transcriptional regulatory elements. This method, extensive to other ligand-modulated transcription factors, might be useful to all transcription factors with slight modifications.
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- Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
- Carr F. E., Wong N. C. Characteristics of a negative thyroid hormone response element. J Biol Chem. 1994 Feb 11;269(6):4175–4179. [PubMed] [Google Scholar]
- Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
- Costa-Giomi M. P., Gaub M. P., Chambon P., Abarzúa P. Characterization of a retinoic acid responsive element isolated by whole genome PCR. Nucleic Acids Res. 1992 Jun 25;20(12):3223–3232. doi: 10.1093/nar/20.12.3223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Danielson P. E., Forss-Petter S., Brow M. A., Calavetta L., Douglass J., Milner R. J., Sutcliffe J. G. p1B15: a cDNA clone of the rat mRNA encoding cyclophilin. DNA. 1988 May;7(4):261–267. doi: 10.1089/dna.1988.7.261. [DOI] [PubMed] [Google Scholar]
- Davis M. M., Cohen D. I., Nielsen E. A., Steinmetz M., Paul W. E., Hood L. Cell-type-specific cDNA probes and the murine I region: the localization and orientation of Ad alpha. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2194–2198. doi: 10.1073/pnas.81.7.2194. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dipple K. M., Qulali M., Ross R. A., Crabb D. W. Effects of thyroxine on the expression of alcohol dehydrogenase in rat liver and kidney. Hepatology. 1993 Apr;17(4):701–706. doi: 10.1002/hep.1840170426. [DOI] [PubMed] [Google Scholar]
- Glass C. K., Holloway J. M. Regulation of gene expression by the thyroid hormone receptor. Biochim Biophys Acta. 1990 Dec 11;1032(2-3):157–176. doi: 10.1016/0304-419x(90)90002-i. [DOI] [PubMed] [Google Scholar]
- Gould A. P., Brookman J. J., Strutt D. I., White R. A. Targets of homeotic gene control in Drosophila. Nature. 1990 Nov 22;348(6299):308–312. doi: 10.1038/348308a0. [DOI] [PubMed] [Google Scholar]
- Inoue S., Kondo S., Hashimoto M., Kondo T., Muramatsu M. Isolation of estrogen receptor-binding sites in human genomic DNA. Nucleic Acids Res. 1991 Aug 11;19(15):4091–4096. doi: 10.1093/nar/19.15.4091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inoue S., Orimo A., Hosoi T., Kondo S., Toyoshima H., Kondo T., Ikegami A., Ouchi Y., Orimo H., Muramatsu M. Genomic binding-site cloning reveals an estrogen-responsive gene that encodes a RING finger protein. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11117–11121. doi: 10.1073/pnas.90.23.11117. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kinzler K. W., Vogelstein B. The GLI gene encodes a nuclear protein which binds specific sequences in the human genome. Mol Cell Biol. 1990 Feb;10(2):634–642. doi: 10.1128/mcb.10.2.634. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kinzler K. W., Vogelstein B. Whole genome PCR: application to the identification of sequences bound by gene regulatory proteins. Nucleic Acids Res. 1989 May 25;17(10):3645–3653. doi: 10.1093/nar/17.10.3645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuhn N. J., Woodworth-Gutai M., Gross K. W., Held W. A. Subfamilies of the mouse major urinary protein (MUP) multi-gene family: sequence analysis of cDNA clones and differential regulation in the liver. Nucleic Acids Res. 1984 Aug 10;12(15):6073–6090. doi: 10.1093/nar/12.15.6073. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liang P., Pardee A. B. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science. 1992 Aug 14;257(5072):967–971. doi: 10.1126/science.1354393. [DOI] [PubMed] [Google Scholar]
- Liaw C. W., Towle H. C. Characterization of a thyroid hormone-responsive gene from rat. J Biol Chem. 1984 Jun 10;259(11):7253–7260. [PubMed] [Google Scholar]
- När A. M., Boutin J. M., Lipkin S. M., Yu V. C., Holloway J. M., Glass C. K., Rosenfeld M. G. The orientation and spacing of core DNA-binding motifs dictate selective transcriptional responses to three nuclear receptors. Cell. 1991 Jun 28;65(7):1267–1279. doi: 10.1016/0092-8674(91)90021-p. [DOI] [PubMed] [Google Scholar]
- Raynal F., Michot B., Bachellerie J. P. Complete nucleotide sequence of mouse 18 S rRNA gene: comparison with other available homologs. FEBS Lett. 1984 Feb 27;167(2):263–268. doi: 10.1016/0014-5793(84)80139-8. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scott M. R., Westphal K. H., Rigby P. W. Activation of mouse genes in transformed cells. Cell. 1983 Sep;34(2):557–567. doi: 10.1016/0092-8674(83)90388-4. [DOI] [PubMed] [Google Scholar]
- Seelig S., Liaw C., Towle H. C., Oppenheimer J. H. Thyroid hormone attenuates and augments hepatic gene expression at a pretranslational level. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4733–4737. doi: 10.1073/pnas.78.8.4733. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shi Y. B., Brown D. D. The earliest changes in gene expression in tadpole intestine induced by thyroid hormone. J Biol Chem. 1993 Sep 25;268(27):20312–20317. [PubMed] [Google Scholar]
- Umesono K., Murakami K. K., Thompson C. C., Evans R. M. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991 Jun 28;65(7):1255–1266. doi: 10.1016/0092-8674(91)90020-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang Z., Brown D. D. A gene expression screen. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11505–11509. doi: 10.1073/pnas.88.24.11505. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinberger C., Thompson C. C., Ong E. S., Lebo R., Gruol D. J., Evans R. M. The c-erb-A gene encodes a thyroid hormone receptor. Nature. 1986 Dec 18;324(6098):641–646. doi: 10.1038/324641a0. [DOI] [PubMed] [Google Scholar]
- Wong G., Itakura T., Kawajiri K., Skow L., Negishi M. Gene family of male-specific testosterone 16 alpha-hydroxylase (C-P-450(16 alpha)) in mice. Organization, differential regulation, and chromosome localization. J Biol Chem. 1989 Feb 15;264(5):2920–2927. [PubMed] [Google Scholar]
- Wright W. E., Funk W. D. CASTing for multicomponent DNA-binding complexes. Trends Biochem Sci. 1993 Mar;18(3):77–80. doi: 10.1016/0968-0004(93)90156-h. [DOI] [PubMed] [Google Scholar]
- el-Deiry W. S., Kern S. E., Pietenpol J. A., Kinzler K. W., Vogelstein B. Definition of a consensus binding site for p53. Nat Genet. 1992 Apr;1(1):45–49. doi: 10.1038/ng0492-45. [DOI] [PubMed] [Google Scholar]



