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. 1995 Aug 11;23(15):2815–2822. doi: 10.1093/nar/23.15.2815

A novel human c-sis mRNA species is transcribed from a promoter in c-sis intron 1 and contains the code for an alternative PDGF B-like protein.

R P Dirks 1, C Onnekink 1, H J Jansen 1, A de Jong 1, H P Bloemers 1
PMCID: PMC307116  PMID: 7659502

Abstract

The human platelet-derived growth factor (PDGF) B chain precursor is usually translated from a 3.5 kb c-sis/PDGF B gene transcript. The first exon of the c-sis/gene contains the code for the signal peptide of the PDGF B chain precursor, preceded by a 1 kb long untranslated sequence with potent translation inhibitory activity. In this paper we show that a novel 2.6 kb c-sis mRNA present in the human choriocarcinoma cell line JEG-3 initiates at an alternative exon 1, which we refer to as exon 1a. The 90 bp long exon 1a is located in the center of the first intron of the gene. It coincides with a very pronounced DNase-I-hypersensitive site and is preceded by a functional promoter. Of the three ATG codons present in exon 1a, the third one perfectly matches the criteria of a consensus start codon. It initiates an open reading frame that is continuous with the code for the PDGF B chain precursor but lacks the code for a signal peptide. We conclude that this novel 2.6 kb c-sis mRNA species lacks the strong translation inhibitory potential of the regular exon 1 and contains the code for a PDGF B-like protein that may be targeted to the cell nucleus.

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Selected References

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

  1. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  2. Barrett T. B., Gajdusek C. M., Schwartz S. M., McDougall J. K., Benditt E. P. Expression of the sis gene by endothelial cells in culture and in vivo. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6772–6774. doi: 10.1073/pnas.81.21.6772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bronzert D. A., Pantazis P., Antoniades H. N., Kasid A., Davidson N., Dickson R. B., Lippman M. E. Synthesis and secretion of platelet-derived growth factor by human breast cancer cell lines. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5763–5767. doi: 10.1073/pnas.84.16.5763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bywater M., Rorsman F., Bongcam-Rudloff E., Mark G., Hammacher A., Heldin C. H., Westermark B., Betsholtz C. Expression of recombinant platelet-derived growth factor A- and B-chain homodimers in rat-1 cells and human fibroblasts reveals differences in protein processing and autocrine effects. Mol Cell Biol. 1988 Jul;8(7):2753–2762. doi: 10.1128/mcb.8.7.2753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chiu I. M., Reddy E. P., Givol D., Robbins K. C., Tronick S. R., Aaronson S. A. Nucleotide sequence analysis identifies the human c-sis proto-oncogene as a structural gene for platelet-derived growth factor. Cell. 1984 May;37(1):123–129. doi: 10.1016/0092-8674(84)90307-6. [DOI] [PubMed] [Google Scholar]
  6. Clarke M. F., Westin E., Schmidt D., Josephs S. F., Ratner L., Wong-Staal F., Gallo R. C., Reitz M. S., Jr Transformation of NIH 3T3 cells by a human c-sis cDNA clone. 1984 Mar 29-Apr 4Nature. 308(5958):464–467. doi: 10.1038/308464a0. [DOI] [PubMed] [Google Scholar]
  7. Dalla-Favera R., Gallo R. C., Giallongo A., Croce C. M. Chromosomal localization of the human homolog (c-sis) of the simian sarcoma virus onc gene. Science. 1982 Nov 12;218(4573):686–688. doi: 10.1126/science.6291150. [DOI] [PubMed] [Google Scholar]
  8. Dirks R. P., Jansen H. J., Gerritsma J., Onnekink C., Bloemers H. P. Localization and functional analysis of DNase-I-hypersensitive sites in the human c-sis/PDGF-B gene transcription unit and its flanking regions. Eur J Biochem. 1993 Feb 1;211(3):509–519. doi: 10.1111/j.1432-1033.1993.tb17577.x. [DOI] [PubMed] [Google Scholar]
  9. Dirks R. P., Jansen H. J., Onnekink C., De Jonge R. J., Bloemers H. P. DNase-I-hypersensitive sites located far upstream of the human c-sis/PDGF-B gene comap with transcriptional enhancers and a silencer and are preceded by (part of) a new transcription unit. Eur J Biochem. 1993 Sep 1;216(2):487–495. doi: 10.1111/j.1432-1033.1993.tb18167.x. [DOI] [PubMed] [Google Scholar]
  10. Doolittle R. F., Hunkapiller M. W., Hood L. E., Devare S. G., Robbins K. C., Aaronson S. A., Antoniades H. N. Simian sarcoma virus onc gene, v-sis, is derived from the gene (or genes) encoding a platelet-derived growth factor. Science. 1983 Jul 15;221(4607):275–277. doi: 10.1126/science.6304883. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Edwards J. B., Delort J., Mallet J. Oligodeoxyribonucleotide ligation to single-stranded cDNAs: a new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification. Nucleic Acids Res. 1991 Oct 11;19(19):5227–5232. doi: 10.1093/nar/19.19.5227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eva A., Robbins K. C., Andersen P. R., Srinivasan A., Tronick S. R., Reddy E. P., Ellmore N. W., Galen A. T., Lautenberger J. A., Papas T. S. Cellular genes analogous to retroviral onc genes are transcribed in human tumour cells. Nature. 1982 Jan 14;295(5845):116–119. doi: 10.1038/295116a0. [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. Fen Z., Daniel T. O. 5' untranslated sequences determine degradative pathway for alternate PDGF B/c-sis mRNA's. Oncogene. 1991 Jun;6(6):953–959. [PubMed] [Google Scholar]
  16. Franklin G. C., Donovan M., Adam G. I., Holmgren L., Pfeifer-Ohlsson S., Ohlsson R. Expression of the human PDGF-B gene is regulated by both positively and negatively acting cell type-specific regulatory elements located in the first intron. EMBO J. 1991 Jun;10(6):1365–1373. doi: 10.1002/j.1460-2075.1991.tb07656.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gazit A., Igarashi H., Chiu I. M., Srinivasan A., Yaniv A., Tronick S. R., Robbins K. C., Aaronson S. A. Expression of the normal human sis/PDGF-2 coding sequence induces cellular transformation. Cell. 1984 Nov;39(1):89–97. doi: 10.1016/0092-8674(84)90194-6. [DOI] [PubMed] [Google Scholar]
  18. Gladwin A. M., Carrier M. J., Beesley J. E., Lelchuk R., Hancock V., Martin J. F. Identification of mRNA for PDGF B-chain in human megakaryocytes isolated using a novel immunomagnetic separation method. Br J Haematol. 1990 Nov;76(3):333–339. doi: 10.1111/j.1365-2141.1990.tb06364.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Goustin A. S., Betsholtz C., Pfeifer-Ohlsson S., Persson H., Rydnert J., Bywater M., Holmgren G., Heldin C. H., Westermark B., Ohlsson R. Coexpression of the sis and myc proto-oncogenes in developing human placenta suggests autocrine control of trophoblast growth. Cell. 1985 May;41(1):301–312. doi: 10.1016/0092-8674(85)90083-2. [DOI] [PubMed] [Google Scholar]
  21. Graves D. T., Owen A. J., Barth R. K., Tempst P., Winoto A., Fors L., Hood L. E., Antoniades H. N. Detection of c-sis transcripts and synthesis of PDGF-like proteins by human osteosarcoma cells. Science. 1984 Nov 23;226(4677):972–974. doi: 10.1126/science.6209798. [DOI] [PubMed] [Google Scholar]
  22. Graves D. T., Owen A. J., Williams S. R., Antoniades H. N. Identification of processing events in the synthesis of platelet-derived growth factor-like proteins by human osteosarcoma cells. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4636–4640. doi: 10.1073/pnas.83.13.4636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gross D. S., Garrard W. T. Nuclease hypersensitive sites in chromatin. Annu Rev Biochem. 1988;57:159–197. doi: 10.1146/annurev.bi.57.070188.001111. [DOI] [PubMed] [Google Scholar]
  24. Hannink M., Donoghue D. J. Biosynthesis of the v-sis gene product: signal sequence cleavage, glycosylation, and proteolytic processing. Mol Cell Biol. 1986 Apr;6(4):1343–1348. doi: 10.1128/mcb.6.4.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Igarashi H., Rao C. D., Siroff M., Leal F., Robbins K. C., Aaronson S. A. Detection of PDGF-2 homodimers in human tumor cells. Oncogene. 1987 Mar;1(1):79–85. [PubMed] [Google Scholar]
  26. Johnsson A., Heldin C. H., Wasteson A., Westermark B., Deuel T. F., Huang J. S., Seeburg P. H., Gray A., Ullrich A., Scrace G. The c-sis gene encodes a precursor of the B chain of platelet-derived growth factor. EMBO J. 1984 May;3(5):921–928. doi: 10.1002/j.1460-2075.1984.tb01908.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Josephs S. F., Guo C., Ratner L., Wong-Staal F. Human-proto-oncogene nucleotide sequences corresponding to the transforming region of simian sarcoma virus. Science. 1984 Feb 3;223(4635):487–491. doi: 10.1126/science.6318322. [DOI] [PubMed] [Google Scholar]
  28. Josephs S. F., Ratner L., Clarke M. F., Westin E. H., Reitz M. S., Wong-Staal F. Transforming potential of human c-sis nucleotide sequences encoding platelet-derived growth factor. Science. 1984 Aug 10;225(4662):636–639. doi: 10.1126/science.6740330. [DOI] [PubMed] [Google Scholar]
  29. Kaetzel D. M., Coyne D. W., Fenstermaker R. A. Transcriptional control of the platelet-derived growth factor subunit genes. Biofactors. 1993 May;4(2):71–81. [PubMed] [Google Scholar]
  30. Kimura H. Schwannoma-derived growth factor must be transported into the nucleus to exert its mitogenic activity. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2165–2169. doi: 10.1073/pnas.90.6.2165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kozak M. An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol. 1991 Nov;115(4):887–903. doi: 10.1083/jcb.115.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lee B. A., Maher D. W., Hannink M., Donoghue D. J. Identification of a signal for nuclear targeting in platelet-derived-growth-factor-related molecules. Mol Cell Biol. 1987 Oct;7(10):3527–3537. doi: 10.1128/mcb.7.10.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Martinet Y., Bitterman P. B., Mornex J. F., Grotendorst G. R., Martin G. R., Crystal R. G. Activated human monocytes express the c-sis proto-oncogene and release a mediator showing PDGF-like activity. Nature. 1986 Jan 9;319(6049):158–160. doi: 10.1038/319158a0. [DOI] [PubMed] [Google Scholar]
  34. Ostman A., Thyberg J., Westermark B., Heldin C. H. PDGF-AA and PDGF-BB biosynthesis: proprotein processing in the Golgi complex and lysosomal degradation of PDGF-BB retained intracellularly. J Cell Biol. 1992 Aug;118(3):509–519. doi: 10.1083/jcb.118.3.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Press R. D., Samols D., Goldthwait D. A. Expression and stability of c-sis mRNA in human glioblastoma cells. Biochemistry. 1988 Jul 26;27(15):5736–5741. doi: 10.1021/bi00415a051. [DOI] [PubMed] [Google Scholar]
  36. Rao C. D., Igarashi H., Chiu I. M., Robbins K. C., Aaronson S. A. Structure and sequence of the human c-sis/platelet-derived growth factor 2 (SIS/PDGF2) transcriptional unit. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2392–2396. doi: 10.1073/pnas.83.8.2392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rao C. D., Pech M., Robbins K. C., Aaronson S. A. The 5' untranslated sequence of the c-sis/platelet-derived growth factor 2 transcript is a potent translational inhibitor. Mol Cell Biol. 1988 Jan;8(1):284–292. doi: 10.1128/mcb.8.1.284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ratner L. Regulation of expression of the c-sis proto-oncogene. Nucleic Acids Res. 1989 Jun 12;17(11):4101–4115. doi: 10.1093/nar/17.11.4101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ratner L., Thielan B., Collins T. Sequences of the 5' portion of the human c-sis gene: characterization of the transcriptional promoter and regulation of expression of the protein product by 5' untranslated mRNA sequences. Nucleic Acids Res. 1987 Aug 11;15(15):6017–6036. doi: 10.1093/nar/15.15.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ross R., Raines E. W., Bowen-Pope D. F. The biology of platelet-derived growth factor. Cell. 1986 Jul 18;46(2):155–169. doi: 10.1016/0092-8674(86)90733-6. [DOI] [PubMed] [Google Scholar]
  41. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sariban E., Kufe D. Expression of the platelet-derived growth factor 1 and 2 genes in human myeloid cell lines and monocytes. Cancer Res. 1988 Aug 15;48(16):4498–4502. [PubMed] [Google Scholar]
  43. Shimokado K., Raines E. W., Madtes D. K., Barrett T. B., Benditt E. P., Ross R. A significant part of macrophage-derived growth factor consists of at least two forms of PDGF. Cell. 1985 Nov;43(1):277–286. doi: 10.1016/0092-8674(85)90033-9. [DOI] [PubMed] [Google Scholar]
  44. Swan D. C., McBride O. W., Robbins K. C., Keithley D. A., Reddy E. P., Aaronson S. A. Chromosomal mapping of the simian sarcoma virus onc gene analogue in human cells. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4691–4695. doi: 10.1073/pnas.79.15.4691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tsukiyama T., Becker P. B., Wu C. ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature. 1994 Feb 10;367(6463):525–532. doi: 10.1038/367525a0. [DOI] [PubMed] [Google Scholar]
  46. Van den Ouweland A. M., Roebroek A. J., Schalken J. A., Claesen C. A., Bloemers H. P., Van de Ven W. J. Structure and nucleotide sequence of the 5' region of the human and feline c-sis proto-oncogenes. Nucleic Acids Res. 1986 Jan 24;14(2):765–778. doi: 10.1093/nar/14.2.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Van den Ouweland A. M., Van Groningen J. J., Schalken J. A., Van Neck H. W., Bloemers H. P., Van de Ven W. J. Genetic organization of the c-sis transcription unit. Nucleic Acids Res. 1987 Feb 11;15(3):959–970. doi: 10.1093/nar/15.3.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Versnel M. A., Hagemeijer A., Bouts M. J., van der Kwast T. H., Hoogsteden H. C. Expression of c-sis (PDGF B-chain) and PDGF A-chain genes in ten human malignant mesothelioma cell lines derived from primary and metastatic tumors. Oncogene. 1988 Jun;2(6):601–605. [PubMed] [Google Scholar]
  49. Waterfield M. D., Scrace G. T., Whittle N., Stroobant P., Johnsson A., Wasteson A., Westermark B., Heldin C. H., Huang J. S., Deuel T. F. Platelet-derived growth factor is structurally related to the putative transforming protein p28sis of simian sarcoma virus. Nature. 1983 Jul 7;304(5921):35–39. doi: 10.1038/304035a0. [DOI] [PubMed] [Google Scholar]
  50. Westin E. H., Wong-Staal F., Gelmann E. P., Dalla-Favera R., Papas T. S., Lautenberger J. A., Eva A., Reddy E. P., Tronick S. R., Aaronson S. A. Expression of cellular homologues of retroviral onc genes in human hematopoietic cells. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2490–2494. doi: 10.1073/pnas.79.8.2490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wiedłocha A., Falnes P. O., Madshus I. H., Sandvig K., Olsnes S. Dual mode of signal transduction by externally added acidic fibroblast growth factor. Cell. 1994 Mar 25;76(6):1039–1051. doi: 10.1016/0092-8674(94)90381-6. [DOI] [PubMed] [Google Scholar]
  52. van Leen R. W., Kastrop P. M., van Roozendaal K. E., Schoenmakers J. G. Sequence divergence and selection of cap sites in the rat gamma-crystallin gene family. Eur J Biochem. 1986 May 15;157(1):203–208. doi: 10.1111/j.1432-1033.1986.tb09657.x. [DOI] [PubMed] [Google Scholar]
  53. van den Ouweland A. M., Breuer M. L., Steenbergh P. H., Schalken J. A., Bloemers H. P., Van de Ven W. J. Comparative analysis of the human and feline c-sis proto-oncogenes. Identification of 5' human c-sis coding sequences that are not homologous to the transforming gene of simian sarcoma virus. Biochim Biophys Acta. 1985 Jun 24;825(2):140–147. doi: 10.1016/0167-4781(85)90097-1. [DOI] [PubMed] [Google Scholar]

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