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. 1992 Nov;12(11):5069–5077. doi: 10.1128/mcb.12.11.5069

The growth-stimulatory effect of simian virus 40 T antigen requires the interaction of insulinlike growth factor 1 with its receptor.

P Porcu 1, A Ferber 1, Z Pietrzkowski 1, C T Roberts 1, M Adamo 1, D LeRoith 1, R Baserga 1
PMCID: PMC360440  PMID: 1406682

Abstract

We have used a plasmid expressing a temperature-sensitive (ts) mutant of simian virus 40 (SV40) T antigen, stably transfected into 3T3 cells, to study the role of insulinlike growth factor 1 (IGF-1) and its receptor in T-antigen-mediated growth. While 3T3 cells do not grow in serum-free medium, in 1% serum, or with the sole addition of either platelet-derived growth factor (PDGF) or IGF-1, cells expressing the tsA T antigen (BALB 58 cells) grow at 34 degrees C in either PDGF or 1% serum but not in IGF-1. At the restrictive temperature (39.6 degrees C), these cells can only grow in 10% serum. We show that BALB 58 cells, at 34 degrees C, have a markedly increased expression of IGF-1 and IGF-1 mRNA and that their growth in 1% serum (at 34 degrees C) is inhibited by an antisense oligodeoxynucleotide to the IGF-1 receptor RNA. When this tsA plasmid is stably transfected into cells constitutively overexpressing the human IGF-1 receptor cDNA, the resulting cell lines show a constitutively phosphorylated IGF-1 receptor and grow in serum-free medium at 34 degrees C (but not at 39.6 degrees C). A functional SV40 T antigen also increases the expression of a plasmid in which the reporter luciferase gene is under the control of a rat IGF-1 promoter. We conclude (i) that the SV40 T antigen induces the expression of IGF-1 and IGF-1 mRNA, at least in part by a transcriptional mechanism, thus altering the growth factors requirements, and (ii) that, in BALB/c3t3 cells, the SV40 T antigen necessitates a functional IGF-1 receptor for its growth-stimulating effect in low serum (or PDGF).

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Selected References

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

  1. Adamo M. L., Ben-Hur H., LeRoith D., Roberts C. T., Jr Transcription initiation in the two leader exons of the rat IGF-I gene occurs from disperse versus localized sites. Biochem Biophys Res Commun. 1991 Apr 30;176(2):887–893. doi: 10.1016/s0006-291x(05)80269-4. [DOI] [PubMed] [Google Scholar]
  2. Adamo M. L., Ben-Hur H., Roberts C. T., Jr, LeRoith D. Regulation of start site usage in the leader exons of the rat insulin-like growth factor-I gene by development, fasting, and diabetes. Mol Endocrinol. 1991 Nov;5(11):1677–1686. doi: 10.1210/mend-5-11-1677. [DOI] [PubMed] [Google Scholar]
  3. Armelin H. A., Armelin M. C., Kelly K., Stewart T., Leder P., Cochran B. H., Stiles C. D. Functional role for c-myc in mitogenic response to platelet-derived growth factor. Nature. 1984 Aug 23;310(5979):655–660. doi: 10.1038/310655a0. [DOI] [PubMed] [Google Scholar]
  4. Beard P., Offord E., Paduwat N., Bruggmann H. SV40 activates transcription from the transferrin receptor promoter by inducing a factor which binds to the CRE/AP-1 recognition sequence. Nucleic Acids Res. 1991 Dec;19(25):7117–7123. doi: 10.1093/nar/19.25.7117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bell G. I., Stempien M. M., Fong N. M., Rall L. B. Sequences of liver cDNAs encoding two different mouse insulin-like growth factor I precursors. Nucleic Acids Res. 1986 Oct 24;14(20):7873–7882. doi: 10.1093/nar/14.20.7873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bourre F., Sarasin A. Targeted mutagenesis of SV40 DNA induced by UV light. Nature. 1983 Sep 1;305(5929):68–70. doi: 10.1038/305068a0. [DOI] [PubMed] [Google Scholar]
  7. Bucci C., Mallucci P., Roberts C. T., Frunzio R., Bruni C. B. Nucleotide sequence of a genomic fragment of the rat IGF-I gene spanning an alternate 5' non coding exon. Nucleic Acids Res. 1989 May 11;17(9):3596–3596. doi: 10.1093/nar/17.9.3596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Campisi J., Pardee A. B. Post-transcriptional control of the onset of DNA synthesis by an insulin-like growth factor. Mol Cell Biol. 1984 Sep;4(9):1807–1814. doi: 10.1128/mcb.4.9.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Clemmons D. R. Multiple hormones stimulate the production of somatomedin by cultured human fibroblasts. J Clin Endocrinol Metab. 1984 May;58(5):850–856. doi: 10.1210/jcem-58-5-850. [DOI] [PubMed] [Google Scholar]
  11. Clemmons D. R., Shaw D. S. Variables controlling somatomedin production by cultured human fibroblasts. J Cell Physiol. 1983 May;115(2):137–142. doi: 10.1002/jcp.1041150206. [DOI] [PubMed] [Google Scholar]
  12. Deppert W., Kurth M., Graessmann M., Graessmann A., Knippschild U. Altered phosphorylation at specific sites confers a mutant phenotype to SV40 wild-type large T antigen in a flat revertant of SV40-transformed cells. Oncogene. 1991 Oct;6(10):1931–1938. [PubMed] [Google Scholar]
  13. Estrov Z., Meir R., Barak Y., Zaizov R., Zadik Z. Human growth hormone and insulin-like growth factor-1 enhance the proliferation of human leukemic blasts. J Clin Oncol. 1991 Mar;9(3):394–399. doi: 10.1200/JCO.1991.9.3.394. [DOI] [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Gai X. X., Rizzo M. G., Lee J., Ullrich A., Baserga R. Abrogation of the requirements for added growth factors in 3T3 cells constitutively expressing the p53 and IGF-1 genes. Oncogene Res. 1988;3(4):377–386. [PubMed] [Google Scholar]
  16. Goldring M. B., Goldring S. R. Cytokines and cell growth control. Crit Rev Eukaryot Gene Expr. 1991;1(4):301–326. [PubMed] [Google Scholar]
  17. Gritz L., Davies J. Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene. 1983 Nov;25(2-3):179–188. doi: 10.1016/0378-1119(83)90223-8. [DOI] [PubMed] [Google Scholar]
  18. Hizuka N., Sukegawa I., Takano K., Asakawa K., Horikawa R., Tsushima T., Shizume K. Characterization of insulin-like growth factor I receptors on human erythroleukemia cell line (K-562 cells). Endocrinol Jpn. 1987 Feb;34(1):81–88. doi: 10.1507/endocrj1954.34.81. [DOI] [PubMed] [Google Scholar]
  19. Janicot M., Flores-Riveros J. R., Lane M. D. The insulin-like growth factor 1 (IGF-1) receptor is responsible for mediating the effects of insulin, IGF-1, and IGF-2 in Xenopus laevis oocytes. J Biol Chem. 1991 May 25;266(15):9382–9391. [PubMed] [Google Scholar]
  20. Jat P. S., Sharp P. A. Cell lines established by a temperature-sensitive simian virus 40 large-T-antigen gene are growth restricted at the nonpermissive temperature. Mol Cell Biol. 1989 Apr;9(4):1672–1681. doi: 10.1128/mcb.9.4.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kaczmarek L., Hyland J. K., Watt R., Rosenberg M., Baserga R. Microinjected c-myc as a competence factor. Science. 1985 Jun 14;228(4705):1313–1315. doi: 10.1126/science.4001943. [DOI] [PubMed] [Google Scholar]
  22. Kaleko M., Rutter W. J., Miller A. D. Overexpression of the human insulinlike growth factor I receptor promotes ligand-dependent neoplastic transformation. Mol Cell Biol. 1990 Feb;10(2):464–473. doi: 10.1128/mcb.10.2.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Koniecki J., Nugent P., Kordowska J., Baserga R. Effect of the SV40 T antigen on the posttranscriptional regulation of the proliferating cell nuclear antigen and DNA polymerase-alpha genes. Cancer Res. 1991 Mar 1;51(5):1465–1471. [PubMed] [Google Scholar]
  24. Lammers R., Gray A., Schlessinger J., Ullrich A. Differential signalling potential of insulin- and IGF-1-receptor cytoplasmic domains. EMBO J. 1989 May;8(5):1369–1375. doi: 10.1002/j.1460-2075.1989.tb03517.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Leof E. B., Wharton W., van Wyk J. J., Pledger W. J. Epidermal growth factor (EGF) and somatomedin C regulate G1 progression in competent BALB/c-3T3 cells. Exp Cell Res. 1982 Sep;141(1):107–115. doi: 10.1016/0014-4827(82)90073-8. [DOI] [PubMed] [Google Scholar]
  26. Lin A. Y., Lee A. S. Induction of two genes by glucose starvation in hamster fibroblasts. Proc Natl Acad Sci U S A. 1984 Feb;81(4):988–992. doi: 10.1073/pnas.81.4.988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lipson K. E., Baserga R. Transcriptional activity of the human thymidine kinase gene determined by a method using the polymerase chain reaction and an intron-specific probe. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9774–9777. doi: 10.1073/pnas.86.24.9774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Liu H. T., Baserga R., Mercer W. E. Adenovirus type 2 activates cell cycle-dependent genes that are a subset of those activated by serum. Mol Cell Biol. 1985 Nov;5(11):2936–2942. doi: 10.1128/mcb.5.11.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McCubrey J. A., Steelman L. S., Mayo M. W., Algate P. A., Dellow R. A., Kaleko M. Growth-promoting effects of insulin-like growth factor-1 (IGF-1) on hematopoietic cells: overexpression of introduced IGF-1 receptor abrogates interleukin-3 dependency of murine factor-dependent cells by a ligand-dependent mechanism. Blood. 1991 Aug 15;78(4):921–929. [PubMed] [Google Scholar]
  30. Pekonen F., Partanen S., Mäkinen T., Rutanen E. M. Receptors for epidermal growth factor and insulin-like growth factor I and their relation to steroid receptors in human breast cancer. Cancer Res. 1988 Mar 1;48(5):1343–1347. [PubMed] [Google Scholar]
  31. Pepe M. G., Ginzton N. H., Lee P. D., Hintz R. L., Greenberg P. L. Receptor binding and mitogenic effects of insulin and insulinlike growth factors I and II for human myeloid leukemic cells. J Cell Physiol. 1987 Nov;133(2):219–227. doi: 10.1002/jcp.1041330204. [DOI] [PubMed] [Google Scholar]
  32. Peyrat J. P., Bonneterre J., Beuscart R., Djiane J., Demaille A. Insulin-like growth factor 1 receptors in human breast cancer and their relation to estradiol and progesterone receptors. Cancer Res. 1988 Nov 15;48(22):6429–6433. [PubMed] [Google Scholar]
  33. Pietrzkowski Z., Lammers R., Carpenter G., Soderquist A. M., Limardo M., Phillips P. D., Ullrich A., Baserga R. Constitutive expression of insulin-like growth factor 1 and insulin-like growth factor 1 receptor abrogates all requirements for exogenous growth factors. Cell Growth Differ. 1992 Apr;3(4):199–205. [PubMed] [Google Scholar]
  34. Radna R. L., Caton Y., Jha K. K., Kaplan P., Li G., Traganos F., Ozer H. L. Growth of immortal simian virus 40 tsA-transformed human fibroblasts is temperature dependent. Mol Cell Biol. 1989 Jul;9(7):3093–3096. doi: 10.1128/mcb.9.7.3093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rappolee D. A., Wang A., Mark D., Werb Z. Novel method for studying mRNA phenotypes in single or small numbers of cells. J Cell Biochem. 1989 Jan;39(1):1–11. doi: 10.1002/jcb.240390102. [DOI] [PubMed] [Google Scholar]
  36. Reiss K., Ferber A., Travali S., Porcu P., Phillips P. D., Baserga R. The protooncogene c-myb increases the expression of insulin-like growth factor 1 and insulin-like growth factor 1 receptor messenger RNAs by a transcriptional mechanism. Cancer Res. 1991 Nov 1;51(21):5997–6000. [PubMed] [Google Scholar]
  37. Resnick-Silverman L., Pang Z., Li G., Jha K. K., Ozer H. L. Retinoblastoma protein and simian virus 40-dependent immortalization of human fibroblasts. J Virol. 1991 Jun;65(6):2845–2852. doi: 10.1128/jvi.65.6.2845-2852.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Russell W. E., Van Wyk J. J., Pledger W. J. Inhibition of the mitogenic effects of plasma by a monoclonal antibody to somatomedin C. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2389–2392. doi: 10.1073/pnas.81.8.2389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Scher C. D., Shepard R. C., Antoniades H. N., Stiles C. D. Platelet-derived growth factor and the regulation of the mammalian fibroblast cell cycle. Biochim Biophys Acta. 1979 Aug 10;560(2):217–241. doi: 10.1016/0304-419x(79)90020-9. [DOI] [PubMed] [Google Scholar]
  40. Smith H. S., Scher C. D., Todaro G. J. Induction of cell division in medium lacking serum growth factor by SV40. Virology. 1971 May;44(2):359–370. doi: 10.1016/0042-6822(71)90267-4. [DOI] [PubMed] [Google Scholar]
  41. Stewart A. J., Johnson M. D., May F. E., Westley B. R. Role of insulin-like growth factors and the type I insulin-like growth factor receptor in the estrogen-stimulated proliferation of human breast cancer cells. J Biol Chem. 1990 Dec 5;265(34):21172–21178. [PubMed] [Google Scholar]
  42. Stiles C. D., Capone G. T., Scher C. D., Antoniades H. N., Van Wyk J. J., Pledger W. J. Dual control of cell growth by somatomedins and platelet-derived growth factor. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1279–1283. doi: 10.1073/pnas.76.3.1279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Surmacz E., Nugent P., Pietrzkowski Z., Baserga R. The role of the IGF1 receptor in the regulation of cdc2 mRNA levels in fibroblasts. Exp Cell Res. 1992 Apr;199(2):275–278. doi: 10.1016/0014-4827(92)90435-b. [DOI] [PubMed] [Google Scholar]
  44. Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. doi: 10.1128/jvi.15.3.613-618.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  46. Travali S., Reiss K., Ferber A., Petralia S., Mercer W. E., Calabretta B., Baserga R. Constitutively expressed c-myb abrogates the requirement for insulinlike growth factor 1 in 3T3 fibroblasts. Mol Cell Biol. 1991 Feb;11(2):731–736. doi: 10.1128/mcb.11.2.731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Ullrich A., Gray A., Tam A. W., Yang-Feng T., Tsubokawa M., Collins C., Henzel W., Le Bon T., Kathuria S., Chen E. Insulin-like growth factor I receptor primary structure: comparison with insulin receptor suggests structural determinants that define functional specificity. EMBO J. 1986 Oct;5(10):2503–2512. doi: 10.1002/j.1460-2075.1986.tb04528.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Werner H., Woloschak M., Adamo M., Shen-Orr Z., Roberts C. T., Jr, LeRoith D. Developmental regulation of the rat insulin-like growth factor I receptor gene. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7451–7455. doi: 10.1073/pnas.86.19.7451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zumstein P., Stiles C. D. Molecular cloning of gene sequences that are regulated by insulin-like growth factor I. J Biol Chem. 1987 Aug 15;262(23):11252–11260. [PubMed] [Google Scholar]
  50. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

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