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. 1990 May;10(5):2035–2040. doi: 10.1128/mcb.10.5.2035

Identification of a novel neuronal C-SRC exon expressed in human brain.

J M Pyper 1, J B Bolen 1
PMCID: PMC360550  PMID: 1691439

Abstract

Neuronal cells are known to express at least two different forms of the C-SRC proto-oncogene as a consequence of alternative splicing events which add an 18-nucleotide exon (the NI exon) between C-SRC exons 3 and 4. Here we report that a second neuronal exon of C-SRC is also present between C-SRC exons 3 and 4. This neuronal exon (the NII exon) of C-SRC was isolated from human adult and fetal brain-derived cDNAs and contains 33 nucleotides capable of encoding 11 amino acids (Gln-Thr-Trp-Phe-Thr-Phe-Arg-Trp-Leu-Gln-Arg). The human NI exon was located approximately 390 nucleotides from the end of C-SRC exon 3, whereas the NII exon was approximately 1,000 nucleotides from the beginning of C-SRC exon 4. Analysis of human brain RNA revealed that the NII exon is utilized primarily in conjunction with the NI exon to yield transcripts capable of encoding C-SRC products possessing 17 additional amino acids. These splicing events, which occur between the NI and NII exons, are predicted to alter the sixth amino acid encoded by the NI exon from an arginine to a serine residue, producing a potentially novel phosphorylation site. Analysis of the different C-SRC RNA transcripts revealed that the level of C-SRC RNA containing both NI and NII exons is similar in adult and fetal brain tissue, whereas the level of C-SRC RNA containing only the NI exon or the nonneuronal form of C-SRC RNAs is significantly higher in fetal brain tissues. These results indicate that the expression and splicing pattern of the C-SRC gene are developmentally regulated in the human brain.

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Selected References

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

  1. Andreadis A., Gallego M. E., Nadal-Ginard B. Generation of protein isoform diversity by alternative splicing: mechanistic and biological implications. Annu Rev Cell Biol. 1987;3:207–242. doi: 10.1146/annurev.cb.03.110187.001231. [DOI] [PubMed] [Google Scholar]
  2. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolen J. B., Rosen N., Israel M. A. Increased pp60c-src tyrosyl kinase activity in human neuroblastomas is associated with amino-terminal tyrosine phosphorylation of the src gene product. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7275–7279. doi: 10.1073/pnas.82.21.7275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Breitbart R. E., Andreadis A., Nadal-Ginard B. Alternative splicing: a ubiquitous mechanism for the generation of multiple protein isoforms from single genes. Annu Rev Biochem. 1987;56:467–495. doi: 10.1146/annurev.bi.56.070187.002343. [DOI] [PubMed] [Google Scholar]
  5. Brugge J. S., Cotton P. C., Queral A. E., Barrett J. N., Nonner D., Keane R. W. Neurones express high levels of a structurally modified, activated form of pp60c-src. Nature. 1985 Aug 8;316(6028):554–557. doi: 10.1038/316554a0. [DOI] [PubMed] [Google Scholar]
  6. Brugge J., Cotton P., Lustig A., Yonemoto W., Lipsich L., Coussens P., Barrett J. N., Nonner D., Keane R. W. Characterization of the altered form of the c-src gene product in neuronal cells. Genes Dev. 1987 May;1(3):287–296. doi: 10.1101/gad.1.3.287. [DOI] [PubMed] [Google Scholar]
  7. Cartwright C. A., Simantov R., Kaplan P. L., Hunter T., Eckhart W. Alterations in pp60c-src accompany differentiation of neurons from rat embryo striatum. Mol Cell Biol. 1987 May;7(5):1830–1840. doi: 10.1128/mcb.7.5.1830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Cooke M. P., Perlmutter R. M. Expression of a novel form of the fyn proto-oncogene in hematopoietic cells. New Biol. 1989 Oct;1(1):66–74. [PubMed] [Google Scholar]
  10. Edwards M. K., McBurney M. W. The concentration of retinoic acid determines the differentiated cell types formed by a teratocarcinoma cell line. Dev Biol. 1983 Jul;98(1):187–191. doi: 10.1016/0012-1606(83)90348-2. [DOI] [PubMed] [Google Scholar]
  11. Fults D. W., Towle A. C., Lauder J. M., Maness P. F. pp60c-src in the developing cerebellum. Mol Cell Biol. 1985 Jan;5(1):27–32. doi: 10.1128/mcb.5.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
  13. Kato J. Y., Takeya T., Grandori C., Iba H., Levy J. B., Hanafusa H. Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein. Mol Cell Biol. 1986 Dec;6(12):4155–4160. doi: 10.1128/mcb.6.12.4155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leff S. E., Rosenfeld M. G., Evans R. M. Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem. 1986;55:1091–1117. doi: 10.1146/annurev.bi.55.070186.005303. [DOI] [PubMed] [Google Scholar]
  15. Levy B. T., Sorge L. K., Meymandi A., Maness P. F. pp60c-src Kinase is in chick and human embryonic tissues. Dev Biol. 1984 Jul;104(1):9–17. doi: 10.1016/0012-1606(84)90031-9. [DOI] [PubMed] [Google Scholar]
  16. Levy J. B., Brugge J. S. Biological and biochemical properties of the c-src+ gene product overexpressed in chicken embryo fibroblasts. Mol Cell Biol. 1989 Aug;9(8):3332–3341. doi: 10.1128/mcb.9.8.3332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levy J. B., Dorai T., Wang L. H., Brugge J. S. The structurally distinct form of pp60c-src detected in neuronal cells is encoded by a unique c-src mRNA. Mol Cell Biol. 1987 Nov;7(11):4142–4145. doi: 10.1128/mcb.7.11.4142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lynch S. A., Brugge J. S., Levine J. M. Induction of altered c-src product during neural differentiation of embryonal carcinoma cells. Science. 1986 Nov 14;234(4778):873–876. doi: 10.1126/science.3095923. [DOI] [PubMed] [Google Scholar]
  19. Maness P. F., Aubry M., Shores C. G., Frame L., Pfenninger K. H. c-src gene product in developing rat brain is enriched in nerve growth cone membranes. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5001–5005. doi: 10.1073/pnas.85.14.5001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Martinez R., Mathey-Prevot B., Bernards A., Baltimore D. Neuronal pp60c-src contains a six-amino acid insertion relative to its non-neuronal counterpart. Science. 1987 Jul 24;237(4813):411–415. doi: 10.1126/science.2440106. [DOI] [PubMed] [Google Scholar]
  21. Mellström K., Bjelfman C., Hammerling U., Påhlman S. Expression of c-src in cultured human neuroblastoma and small-cell lung carcinoma cell lines correlates with neurocrine differentiation. Mol Cell Biol. 1987 Dec;7(12):4178–4184. doi: 10.1128/mcb.7.12.4178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. O'Shaughnessy J., Deseau V., Amini S., Rosen N., Bolen J. B. Analysis of the c-src gene product structure, abundance, and protein kinase activity in human neuroblastoma and glioblastoma cells. Oncogene Res. 1987;2(1):1–18. [PubMed] [Google Scholar]
  23. Potts W. M., Reynolds A. B., Lansing T. J., Parsons J. T. Activation of pp60c-src transforming potential by mutations altering the structure of an amino terminal domain containing residues 90-95. Oncogene Res. 1988;3(4):343–355. [PubMed] [Google Scholar]
  24. Pyper J. M., Bolen J. B. Neuron-specific splicing of C-SRC RNA in human brain. J Neurosci Res. 1989 Sep;24(1):89–96. doi: 10.1002/jnr.490240113. [DOI] [PubMed] [Google Scholar]
  25. Schartl M., Barnekow A. Differential expression of the cellular src gene during vertebrate development. Dev Biol. 1984 Oct;105(2):415–422. doi: 10.1016/0012-1606(84)90298-7. [DOI] [PubMed] [Google Scholar]
  26. Shenoy S., Choi J. K., Bagrodia S., Copeland T. D., Maller J. L., Shalloway D. Purified maturation promoting factor phosphorylates pp60c-src at the sites phosphorylated during fibroblast mitosis. Cell. 1989 Jun 2;57(5):763–774. doi: 10.1016/0092-8674(89)90791-5. [DOI] [PubMed] [Google Scholar]
  27. Sorge J. P., Sorge L. K., Maness P. F. pp60c-src is expressed in human fetal and adult brain. Am J Pathol. 1985 Apr;119(1):151–157. [PMC free article] [PubMed] [Google Scholar]
  28. Sorge L. K., Levy B. T., Maness P. F. pp60c-src is developmentally regulated in the neural retina. Cell. 1984 Feb;36(2):249–257. doi: 10.1016/0092-8674(84)90218-6. [DOI] [PubMed] [Google Scholar]
  29. Sudol M., Alvarez-Buylla A., Hanafusa H. Differential developmental expression of cellular yes and cellular src proteins in cerebellum. Oncogene Res. 1988 May;2(4):345–355. [PubMed] [Google Scholar]
  30. Sutcliffe J. G. mRNA in the mammalian central nervous system. Annu Rev Neurosci. 1988;11:157–198. doi: 10.1146/annurev.ne.11.030188.001105. [DOI] [PubMed] [Google Scholar]
  31. Takeya T., Hanafusa H. Structure and sequence of the cellular gene homologous to the RSV src gene and the mechanism for generating the transforming virus. Cell. 1983 Mar;32(3):881–890. doi: 10.1016/0092-8674(83)90073-9. [DOI] [PubMed] [Google Scholar]
  32. Tanaka A., Gibbs C. P., Arthur R. R., Anderson S. K., Kung H. J., Fujita D. J. DNA sequence encoding the amino-terminal region of the human c-src protein: implications of sequence divergence among src-type kinase oncogenes. Mol Cell Biol. 1987 May;7(5):1978–1983. doi: 10.1128/mcb.7.5.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Veillette A., Bolen J. B. src-related protein tyrosine kinases. Cancer Treat Res. 1989;47:121–142. doi: 10.1007/978-1-4613-1599-5_5. [DOI] [PubMed] [Google Scholar]
  34. Veillette A., Foss F. M., Sausville E. A., Bolen J. B., Rosen N. Expression of the lck tyrosine kinase gene in human colon carcinoma and other non-lymphoid human tumor cell lines. Oncogene Res. 1987 Sep-Oct;1(4):357–374. [PubMed] [Google Scholar]
  35. Veillette A., O'Shaughnessy J., Horak I. D., Israel M. A., Yee D., Rosen N., Fujita D. J., Kung H. J., Biedler J. L., Bolen J. B. Coordinate alteration of pp60c-src abundance and c-src RNA expression in human neuroblastoma variants. Oncogene. 1989 Apr;4(4):421–427. [PubMed] [Google Scholar]
  36. Wang L. H., Iijima S., Dorai T., Lin B. Regulation of the expression of proto-oncogene c-src by alternative RNA splicing in chicken skeletal muscle. Oncogene Res. 1987 Jun;1(1):43–59. [PubMed] [Google Scholar]
  37. Wiestler O. D., Walter G. Developmental expression of two forms of pp60c-src in mouse brain. Mol Cell Biol. 1988 Jan;8(1):502–504. doi: 10.1128/mcb.8.1.502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yang X. M., Walter G. Specific kinase activity and phosphorylation state of pp60c-src from neuroblastomas and fibroblasts. Oncogene. 1988 Sep;3(3):237–244. [PubMed] [Google Scholar]

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