Skip to main content
PMC home page
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
. 1988 Dec;85(23):9076–9080. doi: 10.1073/pnas.85.23.9076

Conditional enhancement of liver-specific gene transcription.

K S Zaret 1, C M DiPersio 1, D A Jackson 1, W J Montigny 1, D L Weinstat 1
PMCID: PMC282666  PMID: 3194409

Abstract

We sought to develop a cell line in which liver-specific transcription could be induced at will, to facilitate the study of factors that cause hepatocyte-specific transcription of the serum albumin gene in mice. We therefore created the H2.35 cell line from mouse hepatocytes infected with a temperature-sensitive strain of simian virus 40. During routine propagation at the permissive temperature, H2.35 cells exhibit extremely low levels of albumin transcription and mRNA. Albumin mRNA increases at least 100-fold when H2.35 cells are cultured at the restrictive temperature and in serum-free medium on a collagen substratum; the two latter conditions maintain the differentiated state of primary hepatocyte cultures. Although a major cause of the mRNA increase is posttranscriptional, the transcription rates of albumin and other liver-specific genes increase significantly. Transient-transfection experiments demonstrated that an induction of transcription is caused by activation of an albumin upstream sequence that was previously shown to enhance liver-specific transcription in transgenic mice. Thus, hepatocyte differentiation appears to be maintained in part by extracellular signals that stimulate the activity of a tissue-specific enhancer element.

Full text

PDF
9077

Images in this article

9077Image
on p.9077
9078Image
on p.9078
9078Image
on p.9078
9078Image
on p.9078
9078Image
on p.9078
9078Image
on p.9078
9078Image
on p.9078
9079Image
on p.9079

Selected References

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

  1. Beale E. G., Chrapkiewicz N. B., Scoble H. A., Metz R. J., Quick D. P., Noble R. L., Donelson J. E., Biemann K., Granner D. K. Rat hepatic cytosolic phosphoenolpyruvate carboxykinase (GTP). Structures of the protein, messenger RNA, and gene. J Biol Chem. 1985 Sep 5;260(19):10748–10760. [PubMed] [Google Scholar]
  2. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  3. Chou J. Y. Temperature-sensitive adult liver cell line dependent on glucocorticoid for differentiation. Mol Cell Biol. 1983 Jun;3(6):1013–1020. doi: 10.1128/mcb.3.6.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clayton D. F., Darnell J. E., Jr Changes in liver-specific compared to common gene transcription during primary culture of mouse hepatocytes. Mol Cell Biol. 1983 Sep;3(9):1552–1561. doi: 10.1128/mcb.3.9.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clayton D. F., Harrelson A. L., Darnell J. E., Jr Dependence of liver-specific transcription on tissue organization. Mol Cell Biol. 1985 Oct;5(10):2623–2632. doi: 10.1128/mcb.5.10.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clayton D. F., Weiss M., Darnell J. E., Jr Liver-specific RNA metabolism in hepatoma cells: variations in transcription rates and mRNA levels. Mol Cell Biol. 1985 Oct;5(10):2633–2641. doi: 10.1128/mcb.5.10.2633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
  8. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
  9. Derman E., Krauter K., Walling L., Weinberger C., Ray M., Darnell J. E., Jr Transcriptional control in the production of liver-specific mRNAs. Cell. 1981 Mar;23(3):731–739. doi: 10.1016/0092-8674(81)90436-0. [DOI] [PubMed] [Google Scholar]
  10. DiMaio D., Nathans D. Regulatory mutants of simian virus 40. Effect of mutations at a T antigen binding site on DNA replication and expression of viral genes. J Mol Biol. 1982 Apr 15;156(3):531–548. doi: 10.1016/0022-2836(82)90265-0. [DOI] [PubMed] [Google Scholar]
  11. Dierks P., van Ooyen A., Mantei N., Weissmann C. DNA sequences preceding the rabbit beta-globin gene are required for formation in mouse L cells of beta-globin RNA with the correct 5' terminus. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1411–1415. doi: 10.1073/pnas.78.3.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Enat R., Jefferson D. M., Ruiz-Opazo N., Gatmaitan Z., Leinwand L. A., Reid L. M. Hepatocyte proliferation in vitro: its dependence on the use of serum-free hormonally defined medium and substrata of extracellular matrix. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1411–1415. doi: 10.1073/pnas.81.5.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fraslin J. M., Kneip B., Vaulont S., Glaise D., Munnich A., Guguen-Guillouzo C. Dependence of hepatocyte-specific gene expression on cell-cell interactions in primary culture. EMBO J. 1985 Oct;4(10):2487–2491. doi: 10.1002/j.1460-2075.1985.tb03960.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Houssaint E. Differentiation of the mouse hepatic primordium. I. An analysis of tissue interactions in hepatocyte differentiation. Cell Differ. 1980 Oct;9(5):269–279. doi: 10.1016/0045-6039(80)90026-3. [DOI] [PubMed] [Google Scholar]
  16. Jefferson D. M., Clayton D. F., Darnell J. E., Jr, Reid L. M. Posttranscriptional modulation of gene expression in cultured rat hepatocytes. Mol Cell Biol. 1984 Sep;4(9):1929–1934. doi: 10.1128/mcb.4.9.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kioussis D., Eiferman F., van de Rijn P., Gorin M. B., Ingram R. S., Tilghman S. M. The evolution of alpha-fetoprotein and albumin. II. The structures of the alpha-fetoprotein and albumin genes in the mouse. J Biol Chem. 1981 Feb 25;256(4):1960–1967. [PubMed] [Google Scholar]
  18. Liu J. K., Bergman Y., Zaret K. S. The mouse albumin promoter and a distal upstream site are simultaneously DNase I hypersensitive in liver chromatin and bind similar liver-abundant factors in vitro. Genes Dev. 1988 May;2(5):528–541. doi: 10.1101/gad.2.5.528. [DOI] [PubMed] [Google Scholar]
  19. McKnight S. L. The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucleic Acids Res. 1980 Dec 20;8(24):5949–5964. doi: 10.1093/nar/8.24.5949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller T. B., Jr, Garnache A., Cruz J. Insulin regulation of glycogen synthase phosphatase in primary cultures of hepatocytes. J Biol Chem. 1984 Oct 25;259(20):12470–12474. [PubMed] [Google Scholar]
  21. Morin O., Normand C. Long-term maintenance of hepatocyte functional activity in co-culture: requirements for sinusoidal endothelial cells and dexamethasone. J Cell Physiol. 1986 Oct;129(1):103–110. doi: 10.1002/jcp.1041290115. [DOI] [PubMed] [Google Scholar]
  22. Noda C., Tomomura M., Nakamura T., Ichihara A. Molecular cloning of DNA complementary to mRNA of rat liver serine dehydratase. Biochem Biophys Res Commun. 1985 Oct 15;132(1):232–239. doi: 10.1016/0006-291x(85)91012-5. [DOI] [PubMed] [Google Scholar]
  23. Pinkert C. A., Ornitz D. M., Brinster R. L., Palmiter R. D. An albumin enhancer located 10 kb upstream functions along with its promoter to direct efficient, liver-specific expression in transgenic mice. Genes Dev. 1987 May;1(3):268–276. doi: 10.1101/gad.1.3.268. [DOI] [PubMed] [Google Scholar]
  24. Powell D. J., Friedman J. M., Oulette A. J., Krauter K. S., Darnell J. E., Jr Transcriptional and post-transcriptional control of specific messenger RNAs in adult and embryonic liver. J Mol Biol. 1984 Oct 15;179(1):21–35. doi: 10.1016/0022-2836(84)90304-8. [DOI] [PubMed] [Google Scholar]
  25. Reid L. M., Jefferson D. M. Culturing hepatocytes and other differentiated cells. Hepatology. 1984 May-Jun;4(3):548–559. doi: 10.1002/hep.1840040332. [DOI] [PubMed] [Google Scholar]
  26. Renkawitz R., Gerbi S. A., Glätzer K. H. Ribosomal DNA of fly Sciara coprophila has a very small and homogeneous repeat unit. Mol Gen Genet. 1979 May 23;173(1):1–13. doi: 10.1007/BF00267685. [DOI] [PubMed] [Google Scholar]
  27. Rio D. C., Clark S. G., Tjian R. A mammalian host-vector system that regulates expression and amplification of transfected genes by temperature induction. Science. 1985 Jan 4;227(4682):23–28. doi: 10.1126/science.2981116. [DOI] [PubMed] [Google Scholar]
  28. Rio D. C., Tjian R. SV40 T antigen binding site mutations that affect autoregulation. Cell. 1983 Apr;32(4):1227–1240. doi: 10.1016/0092-8674(83)90305-7. [DOI] [PubMed] [Google Scholar]
  29. Robins D. M., Paek I., Seeburg P. H., Axel R. Regulated expression of human growth hormone genes in mouse cells. Cell. 1982 Jun;29(2):623–631. doi: 10.1016/0092-8674(82)90178-7. [DOI] [PubMed] [Google Scholar]
  30. Scherer G., Schmid W., Strange C. M., Röwekamp W., Schütz G. Isolation of cDNA clones coding for rat tyrosine aminotransferase. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7205–7208. doi: 10.1073/pnas.79.23.7205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shahan K., Gilmartin M., Derman E. Nucleotide sequences of liver, lachrymal, and submaxillary gland mouse major urinary protein mRNAs: mosaic structure and construction of panels of gene-specific synthetic oligonucleotide probes. Mol Cell Biol. 1987 May;7(5):1938–1946. doi: 10.1128/mcb.7.5.1938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tilghman S. M., Belayew A. Transcriptional control of the murine albumin/alpha-fetoprotein locus during development. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5254–5257. doi: 10.1073/pnas.79.17.5254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Unterman R. D., Lynch K. R., Nakhasi H. L., Dolan K. P., Hamilton J. W., Cohn D. V., Feigelson P. Cloning and sequence of several alpha 2u-globulin cDNAs. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3478–3482. doi: 10.1073/pnas.78.6.3478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Valenzuela P., Quiroga M., Zaldivar J., Rutter W. J., Kirschner M. W., Cleveland D. W. Nucleotide and corresponding amino acid sequences encoded by alpha and beta tubulin mRNAs. Nature. 1981 Feb 19;289(5799):650–655. doi: 10.1038/289650a0. [DOI] [PubMed] [Google Scholar]
  36. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  37. Woodworth C., Secott T., Isom H. C. Transformation of rat hepatocytes by transfection with simian virus 40 DNA to yield proliferating differentiated cells. Cancer Res. 1986 Aug;46(8):4018–4026. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES