Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1994 Aug;94(2):808–814. doi: 10.1172/JCI117400

LAP (NF-IL6) transactivates the collagen alpha 1(I) gene from a 5' regulatory region.

K Houglum 1, M Buck 1, V Adir 1, M Chojkier 1
PMCID: PMC296161  PMID: 8040336

Abstract

Although collagen is known to enhance hepatocyte differentiation and hepatocytes produce collagen in vivo, the transcriptional factors responsible for collagen type I gene expression in hepatic cells are not known. LAP (Liver Activator Protein) is a member of the C/EBP family, which in differentiated hepatocytes contributes to the high levels of liver-specific gene expression. In this study we show that LAP binds to the collagen alpha 1(I) promoter at both reverse CCAAT motifs and activates transcription. Furthermore, an upstream element, collagen element I (-370/-344), which shares homology with the LAP binding cis-element of the albumin promoter (9 of 13 bp) is described. This collagen element I stimulates transcription in both orientations and when placed in front of either a homologous or a heterologous chimeric report construct. These experiments suggest that LAP may be important in the expression of collagen in differentiated hepatocytes through both the promoter and a newly described upstream element.

Full text

PDF
811

Images in this article

809Image
on p.809
810Image
on p.810
810Image
on p.810
811Image
on p.811
811Image
on p.811
812Image
on p.812

Selected References

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

  1. Akira S., Isshiki H., Sugita T., Tanabe O., Kinoshita S., Nishio Y., Nakajima T., Hirano T., Kishimoto T. A nuclear factor for IL-6 expression (NF-IL6) is a member of a C/EBP family. EMBO J. 1990 Jun;9(6):1897–1906. doi: 10.1002/j.1460-2075.1990.tb08316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ben-Ze'ev A., Robinson G. S., Bucher N. L., Farmer S. R. Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2161–2165. doi: 10.1073/pnas.85.7.2161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bissell D. M., Caron J. M., Babiss L. E., Friedman J. M. Transcriptional regulation of the albumin gene in cultured rat hepatocytes. Role of basement-membrane matrix. Mol Biol Med. 1990 Apr;7(2):187–197. [PubMed] [Google Scholar]
  4. Boffini A., Prentki P. Identification of protein binding sites in genomic DNA by two-dimensional gel electrophoresis. Nucleic Acids Res. 1991 Apr 11;19(7):1369–1374. doi: 10.1093/nar/19.7.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bornstein P., McKay J. The first intron of the alpha 1(I) collagen gene contains several transcriptional regulatory elements. J Biol Chem. 1988 Feb 5;263(4):1603–1606. [PubMed] [Google Scholar]
  6. Bornstein P., Sage H. Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol. 1989;37:67–106. doi: 10.1016/s0079-6603(08)60695-9. [DOI] [PubMed] [Google Scholar]
  7. Brenner D. A., Alcorn J. M., Feitelberg S. P., Leffert H. L., Chojkier M. Expression of collagen genes in the liver. Mol Biol Med. 1990 Apr;7(2):105–115. [PubMed] [Google Scholar]
  8. Brenner D. A., Chojkier M. Acetaldehyde increases collagen gene transcription in cultured human fibroblasts. J Biol Chem. 1987 Dec 25;262(36):17690–17695. [PubMed] [Google Scholar]
  9. Buck M., Turler H., Chojkier M. LAP (NF-IL-6), a tissue-specific transcriptional activator, is an inhibitor of hepatoma cell proliferation. EMBO J. 1994 Feb 15;13(4):851–860. doi: 10.1002/j.1460-2075.1994.tb06328.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cao Z., Umek R. M., McKnight S. L. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 1991 Sep;5(9):1538–1552. doi: 10.1101/gad.5.9.1538. [DOI] [PubMed] [Google Scholar]
  11. Chojkier M., Brenner D. A., Leffert H. L. Vasopressin inhibits type-I collagen and albumin gene expression in primary cultures of adult rat hepatocytes. J Biol Chem. 1989 Jun 5;264(16):9583–9591. [PubMed] [Google Scholar]
  12. Chojkier M., Brenner D. A. Therapeutic strategies for hepatic fibrosis. Hepatology. 1988 Jan-Feb;8(1):176–182. doi: 10.1002/hep.1840080132. [DOI] [PubMed] [Google Scholar]
  13. Chojkier M. Hepatocyte collagen production in vivo in normal rats. J Clin Invest. 1986 Aug;78(2):333–339. doi: 10.1172/JCI112581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Chojkier M., Houglum K., Solis-Herruzo J., Brenner D. A. Stimulation of collagen gene expression by ascorbic acid in cultured human fibroblasts. A role for lipid peroxidation? J Biol Chem. 1989 Oct 5;264(28):16957–16962. [PubMed] [Google Scholar]
  15. Clement B., Grimaud J. A., Campion J. P., Deugnier Y., Guillouzo A. Cell types involved in collagen and fibronectin production in normal and fibrotic human liver. Hepatology. 1986 Mar-Apr;6(2):225–234. doi: 10.1002/hep.1840060212. [DOI] [PubMed] [Google Scholar]
  16. Corlu A., Kneip B., Lhadi C., Leray G., Glaise D., Baffet G., Bourel D., Guguen-Guillouzo C. A plasma membrane protein is involved in cell contact-mediated regulation of tissue-specific genes in adult hepatocytes. J Cell Biol. 1991 Oct;115(2):505–515. doi: 10.1083/jcb.115.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Descombes P., Chojkier M., Lichtsteiner S., Falvey E., Schibler U. LAP, a novel member of the C/EBP gene family, encodes a liver-enriched transcriptional activator protein. Genes Dev. 1990 Sep;4(9):1541–1551. doi: 10.1101/gad.4.9.1541. [DOI] [PubMed] [Google Scholar]
  18. Descombes P., Schibler U. A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell. 1991 Nov 1;67(3):569–579. doi: 10.1016/0092-8674(91)90531-3. [DOI] [PubMed] [Google Scholar]
  19. DiPersio C. M., Jackson D. A., Zaret K. S. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol Cell Biol. 1991 Sep;11(9):4405–4414. doi: 10.1128/mcb.11.9.4405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Exton J. H. Role of phosphoinositides in the regulation of liver function. Hepatology. 1988 Jan-Feb;8(1):152–166. doi: 10.1002/hep.1840080129. [DOI] [PubMed] [Google Scholar]
  22. Faber M., Sealy L. Rous sarcoma virus enhancer factor I is a ubiquitous CCAAT transcription factor highly related to CBF and NF-Y. J Biol Chem. 1990 Dec 25;265(36):22243–22254. [PubMed] [Google Scholar]
  23. Friedman A. D., Landschulz W. H., McKnight S. L. CCAAT/enhancer binding protein activates the promoter of the serum albumin gene in cultured hepatoma cells. Genes Dev. 1989 Sep;3(9):1314–1322. doi: 10.1101/gad.3.9.1314. [DOI] [PubMed] [Google Scholar]
  24. Hooft van Huijsduijnen R., Li X. Y., Black D., Matthes H., Benoist C., Mathis D. Co-evolution from yeast to mouse: cDNA cloning of the two NF-Y (CP-1/CBF) subunits. EMBO J. 1990 Oct;9(10):3119–3127. doi: 10.1002/j.1460-2075.1990.tb07509.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Houglum K., Brenner D. A., Chojkier M. d-alpha-tocopherol inhibits collagen alpha 1(I) gene expression in cultured human fibroblasts. Modulation of constitutive collagen gene expression by lipid peroxidation. J Clin Invest. 1991 Jun;87(6):2230–2235. doi: 10.1172/JCI115258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Karsenty G., de Crombrugghe B. Two different negative and one positive regulatory factors interact with a short promoter segment of the alpha 1 (I) collagen gene. J Biol Chem. 1990 Jun 15;265(17):9934–9942. [PubMed] [Google Scholar]
  27. Liska D. J., Slack J. L., Bornstein P. A highly conserved intronic sequence is involved in transcriptional regulation of the alpha 1(I) collagen gene. Cell Regul. 1990 May;1(6):487–498. doi: 10.1091/mbc.1.6.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Maity S. N., Golumbek P. T., Karsenty G., de Crombrugghe B. Selective activation of transcription by a novel CCAAT binding factor. Science. 1988 Jul 29;241(4865):582–585. doi: 10.1126/science.3399893. [DOI] [PubMed] [Google Scholar]
  30. Mueller C. R., Maire P., Schibler U. DBP, a liver-enriched transcriptional activator, is expressed late in ontogeny and its tissue specificity is determined posttranscriptionally. Cell. 1990 Apr 20;61(2):279–291. doi: 10.1016/0092-8674(90)90808-r. [DOI] [PubMed] [Google Scholar]
  31. Nehls M. C., Rippe R. A., Veloz L., Brenner D. A. Transcription factors nuclear factor I and Sp1 interact with the murine collagen alpha 1 (I) promoter. Mol Cell Biol. 1991 Aug;11(8):4065–4073. doi: 10.1128/mcb.11.8.4065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Poli V., Mancini F. P., Cortese R. IL-6DBP, a nuclear protein involved in interleukin-6 signal transduction, defines a new family of leucine zipper proteins related to C/EBP. Cell. 1990 Nov 2;63(3):643–653. doi: 10.1016/0092-8674(90)90459-r. [DOI] [PubMed] [Google Scholar]
  33. Raghow R., Gossage D., Seyer J. M., Kang A. H. Transcriptional regulation of type I collagen genes in cultured fibroblasts by a factor isolated from thioacetamide-induced fibrotic rat liver. J Biol Chem. 1984 Oct 25;259(20):12718–12723. [PubMed] [Google Scholar]
  34. Raghow R., Postlethwaite A. E., Keski-Oja J., Moses H. L., Kang A. H. Transforming growth factor-beta increases steady state levels of type I procollagen and fibronectin messenger RNAs posttranscriptionally in cultured human dermal fibroblasts. J Clin Invest. 1987 Apr;79(4):1285–1288. doi: 10.1172/JCI112950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ravazzolo R., Karsenty G., de Crombrugghe B. A fibroblast-specific factor binds to an upstream negative control element in the promoter of the mouse alpha 1(I) collagen gene. J Biol Chem. 1991 Apr 25;266(12):7382–7387. [PubMed] [Google Scholar]
  36. Rossi P., Karsenty G., Roberts A. B., Roche N. S., Sporn M. B., de Crombrugghe B. A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-beta. Cell. 1988 Feb 12;52(3):405–414. doi: 10.1016/s0092-8674(88)80033-3. [DOI] [PubMed] [Google Scholar]
  37. Rossouw C. M., Vergeer W. P., du Plooy S. J., Bernard M. P., Ramirez F., de Wet W. J. DNA sequences in the first intron of the human pro-alpha 1(I) collagen gene enhance transcription. J Biol Chem. 1987 Nov 5;262(31):15151–15157. [PubMed] [Google Scholar]
  38. Sawada N., Tomomura A., Sattler C. A., Sattler G. L., Kleinman H. K., Pitot H. C. Extracellular matrix components influence DNA synthesis of rat hepatocytes in primary culture. Exp Cell Res. 1986 Dec;167(2):458–470. doi: 10.1016/0014-4827(86)90186-2. [DOI] [PubMed] [Google Scholar]
  39. Slack J. L., Liska D. J., Bornstein P. An upstream regulatory region mediates high-level, tissue-specific expression of the human alpha 1(I) collagen gene in transgenic mice. Mol Cell Biol. 1991 Apr;11(4):2066–2074. doi: 10.1128/mcb.11.4.2066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  41. Trautwein C., Caelles C., van der Geer P., Hunter T., Karin M., Chojkier M. Transactivation by NF-IL6/LAP is enhanced by phosphorylation of its activation domain. Nature. 1993 Aug 5;364(6437):544–547. doi: 10.1038/364544a0. [DOI] [PubMed] [Google Scholar]
  42. Wegner M., Cao Z., Rosenfeld M. G. Calcium-regulated phosphorylation within the leucine zipper of C/EBP beta. Science. 1992 Apr 17;256(5055):370–373. doi: 10.1126/science.256.5055.370. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES