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. 1990 Jul;10(7):3782–3787. doi: 10.1128/mcb.10.7.3782

Extinction of phosphoenolpyruvate carboxykinase gene expression is associated with loss of a specific chromatin-binding protein from a far upstream domain.

Y T Ip 1, R E Fournier 1, R Chalkley 1
PMCID: PMC360833  PMID: 2355923

Abstract

We have analyzed the chromatin structure of the phosphoenolpyruvate carboxykinase (PEPCK) gene in hepatoma x fibroblast hybrids with different extinction phenotypes. These hybrids included a karyotypically complete hybrid in which all liver gene activity was extinguished, a microcell hybrid that contained a single mouse chromosome 11 and in which PEPCK gene activity was decreased but inducible by cyclic AMP, and a segregant line that had lost all mouse chromosomes and in which the PEPCK gene was reexpressed. We found that only in the completely extinguished hybrid was PEPCK chromatin structure radically different from that in the parental hepatoma cells. In this hybrid, there was no evidence of any factors binding to the promoter or to the upstream hypersensitive site at -4800 base pairs. In the other cell lines, even when PEPCK gene transcription was low, the PEPCK chromatin showed characteristic structures typical of a transcriptionally competent gene, with hypersensitive sites at positions previously described. Loss of the upstream hypersensitive site was also shown to be correlated with the absence of a liver-specific protein factor that binds specifically to the upstream region.

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Selected References

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

  1. Chin A. C., Fournier R. E. A genetic analysis of extinction: trans-regulation of 16 liver-specific genes in hepatoma-fibroblast hybrid cells. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1614–1618. doi: 10.1073/pnas.84.6.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chin A. C., Fournier R. E. Tse-2: a trans-dominant extinguisher of albumin gene expression in hepatoma hybrid cells. Mol Cell Biol. 1989 Sep;9(9):3736–3743. doi: 10.1128/mcb.9.9.3736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Costa R. H., Grayson D. R., Darnell J. E., Jr Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol Cell Biol. 1989 Apr;9(4):1415–1425. doi: 10.1128/mcb.9.4.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Costa R. H., Grayson D. R., Xanthopoulos K. G., Darnell J. E., Jr A liver-specific DNA-binding protein recognizes multiple nucleotide sites in regulatory regions of transthyretin, alpha 1-antitrypsin, albumin, and simian virus 40 genes. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3840–3844. doi: 10.1073/pnas.85.11.3840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davidson R. L. Gene expression in somatic cell hybrids. Annu Rev Genet. 1974;8:195–218. doi: 10.1146/annurev.ge.08.120174.001211. [DOI] [PubMed] [Google Scholar]
  6. Ip Y. T., Granner D. K., Chalkley R. Hormonal regulation of phosphoenolpyruvate carboxykinase gene expression is mediated through modulation of an already disrupted chromatin structure. Mol Cell Biol. 1989 Mar;9(3):1289–1297. doi: 10.1128/mcb.9.3.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ip Y. T., Poon D., Stone D., Granner D. K., Chalkley R. Interaction of a liver-specific factor with an enhancer 4.8 kilobases upstream of the phosphoenolpyruvate carboxykinase gene. Mol Cell Biol. 1990 Jul;10(7):3770–3781. doi: 10.1128/mcb.10.7.3770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jones N. C., Rigby P. W., Ziff E. B. Trans-acting protein factors and the regulation of eukaryotic transcription: lessons from studies on DNA tumor viruses. Genes Dev. 1988 Mar;2(3):267–281. doi: 10.1101/gad.2.3.267. [DOI] [PubMed] [Google Scholar]
  9. Killary A. M., Fournier R. E. A genetic analysis of extinction: trans-dominant loci regulate expression of liver-specific traits in hepatoma hybrid cells. Cell. 1984 Sep;38(2):523–534. doi: 10.1016/0092-8674(84)90507-5. [DOI] [PubMed] [Google Scholar]
  10. Lem J., Chin A. C., Thayer M. J., Leach R. J., Fournier R. E. Coordinate regulation of two genes encoding gluconeogenic enzymes by the trans-dominant locus Tse-1. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7302–7306. doi: 10.1073/pnas.85.19.7302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Loose D. S., Shaw P. A., Krauter K. S., Robinson C., Englard S., Hanson R. W., Gluecksohn-Waelsch S. Trans regulation of the phosphoenolpyruvate carboxykinase (GTP) gene, identified by deletions in chromosome 7 of the mouse. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5184–5188. doi: 10.1073/pnas.83.14.5184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maniatis T., Goodbourn S., Fischer J. A. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. doi: 10.1126/science.3296191. [DOI] [PubMed] [Google Scholar]
  13. Quinn P. G., Wong T. W., Magnuson M. A., Shabb J. B., Granner D. K. Identification of basal and cyclic AMP regulatory elements in the promoter of the phosphoenolpyruvate carboxykinase gene. Mol Cell Biol. 1988 Aug;8(8):3467–3475. doi: 10.1128/mcb.8.8.3467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sasaki K., Cripe T. P., Koch S. R., Andreone T. L., Petersen D. D., Beale E. G., Granner D. K. Multihormonal regulation of phosphoenolpyruvate carboxykinase gene transcription. The dominant role of insulin. J Biol Chem. 1984 Dec 25;259(24):15242–15251. [PubMed] [Google Scholar]
  15. Thayer M. J., Fournier R. E. Hormonal regulation of TSE1-repressed genes: evidence for multiple genetic controls in extinction. Mol Cell Biol. 1989 Jul;9(7):2837–2846. doi: 10.1128/mcb.9.7.2837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Weiss M. C., Sparkes R. S., Bertolotti R. Expression of differentiated functions in hepatoma cell hybrids: IX extinction and reexpression of liver-specific enzymes in rat hepatoma-Chinese hamster fibroblast hybrids. Somatic Cell Genet. 1975 Jan;1(1):27–40. doi: 10.1007/BF01538730. [DOI] [PubMed] [Google Scholar]

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