Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Nov;14(11):7182–7194. doi: 10.1128/mcb.14.11.7182

Immortalization-susceptible elements and their binding factors mediate rejuvenation of regulation of the type I collagenase gene in simian virus 40 large T antigen-transformed immortal human fibroblasts.

S Imai 1, T Fujino 1, S Nishibayashi 1, T Manabe 1, T Takano 1
PMCID: PMC359252  PMID: 7935433

Abstract

Dramatic changes occur in expression of the type I collagenase gene during the process of immortalization in simian virus 40 large T antigen-transformed human fibroblasts (S. Imai and T. Takano, Biochem. Biophys. Res. Commun. 189:148-153, 1992). From transient transfection assays, it was determined that these changes involved the functions of two immortalization-susceptible cis-acting elements, ISE1 and ISE2, located in a 100-bp region about 1.7 kb upstream. The profiles of binding of an activator, Proserpine, to the enhancer ISE1 were similar in the extracts of young, senescent preimmortalized and immortalized cells. ISE2 contained both negative and positive regulatory elements located adjacent to each other. The positive regulatory element consisted of a tandem array of putative Ets family- and AP-1-binding sites. An activator, Pluto, interacted with this positive regulatory element and had an AP-1-related component as a complex. The binding activity of Pluto was predominantly detected only in the extract from senescent preimmortalized cells. In contrast, a repressor, Orpheus, which bound to the ATG-rich negative regulatory element of ISE2, was prominently detected in extracts from both young preimmortalized and immortalized cells and appeared to suppress transcription in an orientation-dependent manner. Thus, the interplay of Pluto and Orpheus was suggested to be crucial for regulation of the collagenase gene accompanying in vitro aging and immortalization. Proserpine seemed to interact with Pluto to mediate strong expression of the collagenase gene in cellular senescence. On the basis of these results, we propose a model for regulation of the collagenase gene during in vitro aging and immortalization.

Full text

PDF
7182

Images in this article

7184Image
on p.7184
7189Image
on p.7189
7189Image
on p.7189
7190Image
on p.7190
7191Image
on p.7191

Selected References

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

  1. Angel P., Baumann I., Stein B., Delius H., Rahmsdorf H. J., Herrlich P. 12-O-tetradecanoyl-phorbol-13-acetate induction of the human collagenase gene is mediated by an inducible enhancer element located in the 5'-flanking region. Mol Cell Biol. 1987 Jun;7(6):2256–2266. doi: 10.1128/mcb.7.6.2256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  3. Bauer E. A., Silverman N., Busiek D. F., Kronberger A., Deuel T. F. Diminished response of Werner's syndrome fibroblasts to growth factors PDGF and FGF. Science. 1986 Dec 5;234(4781):1240–1243. doi: 10.1126/science.3022382. [DOI] [PubMed] [Google Scholar]
  4. Dickinson L. A., Joh T., Kohwi Y., Kohwi-Shigematsu T. A tissue-specific MAR/SAR DNA-binding protein with unusual binding site recognition. Cell. 1992 Aug 21;70(4):631–645. doi: 10.1016/0092-8674(92)90432-c. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Edlund T., Walker M. D., Barr P. J., Rutter W. J. Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5' flanking elements. Science. 1985 Nov 22;230(4728):912–916. doi: 10.1126/science.3904002. [DOI] [PubMed] [Google Scholar]
  7. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HAYFLICK L., MOORHEAD P. S. The serial cultivation of human diploid cell strains. Exp Cell Res. 1961 Dec;25:585–621. doi: 10.1016/0014-4827(61)90192-6. [DOI] [PubMed] [Google Scholar]
  9. Hara E., Tsurui H., Shinozaki A., Nakada S., Oda K. Cooperative effect of antisense-Rb and antisense-p53 oligomers on the extension of life span in human diploid fibroblasts, TIG-1. Biochem Biophys Res Commun. 1991 Aug 30;179(1):528–534. doi: 10.1016/0006-291x(91)91403-y. [DOI] [PubMed] [Google Scholar]
  10. Hensler P. J., Annab L. A., Barrett J. C., Pereira-Smith O. M. A gene involved in control of human cellular senescence on human chromosome 1q. Mol Cell Biol. 1994 Apr;14(4):2291–2297. doi: 10.1128/mcb.14.4.2291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  12. Imai S., Saito F., Ikeuchi T., Segawa K., Takano T. Escape from in vitro aging in SV40 large T antigen-transformed human diploid cells: a key event responsible for immortalization occurs during crisis. Mech Ageing Dev. 1993 Jun;69(1-2):149–158. doi: 10.1016/0047-6374(93)90079-7. [DOI] [PubMed] [Google Scholar]
  13. Imai S., Takano T. Loss of collagenase gene expression in immortalized clones of SV40 T antigen-transformed human diploid fibroblasts. Biochem Biophys Res Commun. 1992 Nov 30;189(1):148–153. doi: 10.1016/0006-291x(92)91537-z. [DOI] [PubMed] [Google Scholar]
  14. Jain J., McCaffrey P. G., Valge-Archer V. E., Rao A. Nuclear factor of activated T cells contains Fos and Jun. Nature. 1992 Apr 30;356(6372):801–804. doi: 10.1038/356801a0. [DOI] [PubMed] [Google Scholar]
  15. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  16. Lee W., Mitchell P., Tjian R. Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell. 1987 Jun 19;49(6):741–752. doi: 10.1016/0092-8674(87)90612-x. [DOI] [PubMed] [Google Scholar]
  17. Linder S., Marshall H. Immortalization of primary cells by DNA tumor viruses. Exp Cell Res. 1990 Nov;191(1):1–7. doi: 10.1016/0014-4827(90)90027-8. [DOI] [PubMed] [Google Scholar]
  18. Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Masuda E. S., Tokumitsu H., Tsuboi A., Shlomai J., Hung P., Arai K., Arai N. The granulocyte-macrophage colony-stimulating factor promoter cis-acting element CLE0 mediates induction signals in T cells and is recognized by factors related to AP1 and NFAT. Mol Cell Biol. 1993 Dec;13(12):7399–7407. doi: 10.1128/mcb.13.12.7399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McKnight S. L., Kingsbury R. Transcriptional control signals of a eukaryotic protein-coding gene. Science. 1982 Jul 23;217(4557):316–324. doi: 10.1126/science.6283634. [DOI] [PubMed] [Google Scholar]
  21. Millis A. J., Hoyle M., McCue H. M., Martini H. Differential expression of metalloproteinase and tissue inhibitor of metalloproteinase genes in aged human fibroblasts. Exp Cell Res. 1992 Aug;201(2):373–379. doi: 10.1016/0014-4827(92)90286-h. [DOI] [PubMed] [Google Scholar]
  22. Ning Y., Weber J. L., Killary A. M., Ledbetter D. H., Smith J. R., Pereira-Smith O. M. Genetic analysis of indefinite division in human cells: evidence for a cell senescence-related gene(s) on human chromosome 4. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5635–5639. doi: 10.1073/pnas.88.13.5635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nishina H., Sato H., Suzuki T., Sato M., Iba H. Isolation and characterization of fra-2, an additional member of the fos gene family. Proc Natl Acad Sci U S A. 1990 May;87(9):3619–3623. doi: 10.1073/pnas.87.9.3619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ogata T., Ayusawa D., Namba M., Takahashi E., Oshimura M., Oishi M. Chromosome 7 suppresses indefinite division of nontumorigenic immortalized human fibroblast cell lines KMST-6 and SUSM-1. Mol Cell Biol. 1993 Oct;13(10):6036–6043. doi: 10.1128/mcb.13.10.6036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pereira-Smith O. M., Smith J. R. Evidence for the recessive nature of cellular immortality. Science. 1983 Sep 2;221(4614):964–966. doi: 10.1126/science.6879195. [DOI] [PubMed] [Google Scholar]
  26. Pereira-Smith O. M., Smith J. R. Genetic analysis of indefinite division in human cells: identification of four complementation groups. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6042–6046. doi: 10.1073/pnas.85.16.6042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shay J. W., Pereira-Smith O. M., Wright W. E. A role for both RB and p53 in the regulation of human cellular senescence. Exp Cell Res. 1991 Sep;196(1):33–39. doi: 10.1016/0014-4827(91)90453-2. [DOI] [PubMed] [Google Scholar]
  28. Shay J. W., Wright W. E., Werbin H. Defining the molecular mechanisms of human cell immortalization. Biochim Biophys Acta. 1991 Apr 16;1072(1):1–7. doi: 10.1016/0304-419x(91)90003-4. [DOI] [PubMed] [Google Scholar]
  29. Sottile J., Mann D. M., Diemer V., Millis A. J. Regulation of collagenase and collagenase mRNA production in early- and late-passage human diploid fibroblasts. J Cell Physiol. 1989 Feb;138(2):281–290. doi: 10.1002/jcp.1041380209. [DOI] [PubMed] [Google Scholar]
  30. Sugawara O., Oshimura M., Koi M., Annab L. A., Barrett J. C. Induction of cellular senescence in immortalized cells by human chromosome 1. Science. 1990 Feb 9;247(4943):707–710. doi: 10.1126/science.2300822. [DOI] [PubMed] [Google Scholar]
  31. Suzuki T., Hashimoto Y., Okuno H., Sato H., Nishina H., Iba H. High-level expression of human c-jun gene causes cellular transformation of chicken embryo fibroblasts. Jpn J Cancer Res. 1991 Jan;82(1):58–64. doi: 10.1111/j.1349-7006.1991.tb01746.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thompson C. B., Wang C. Y., Ho I. C., Bohjanen P. R., Petryniak B., June C. H., Miesfeldt S., Zhang L., Nabel G. J., Karpinski B. cis-acting sequences required for inducible interleukin-2 enhancer function bind a novel Ets-related protein, Elf-1. Mol Cell Biol. 1992 Mar;12(3):1043–1053. doi: 10.1128/mcb.12.3.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Thweatt R., Goldstein S. Werner syndrome and biological ageing: a molecular genetic hypothesis. Bioessays. 1993 Jun;15(6):421–426. doi: 10.1002/bies.950150609. [DOI] [PubMed] [Google Scholar]
  34. Wasylyk B., Hahn S. L., Giovane A. The Ets family of transcription factors. Eur J Biochem. 1993 Jan 15;211(1-2):7–18. doi: 10.1007/978-3-642-78757-7_2. [DOI] [PubMed] [Google Scholar]
  35. West M. D., Pereira-Smith O. M., Smith J. R. Replicative senescence of human skin fibroblasts correlates with a loss of regulation and overexpression of collagenase activity. Exp Cell Res. 1989 Sep;184(1):138–147. doi: 10.1016/0014-4827(89)90372-8. [DOI] [PubMed] [Google Scholar]
  36. Wright W. E., Pereira-Smith O. M., Shay J. W. Reversible cellular senescence: implications for immortalization of normal human diploid fibroblasts. Mol Cell Biol. 1989 Jul;9(7):3088–3092. doi: 10.1128/mcb.9.7.3088. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES