Relationship between c-src tyrosine kinase activity and the control of glucose transporter gene expression.

It has been shown previously that the rate of glucose transport in fibroblasts is accelerated by oncoproteins such as v-src, ras, and the transforming protein of feline sarcoma virus. This induction of glucose transport is associated with, and presumably caused by, induction of Hep-G2/rat brain glucose transporter gene expression. To determine the mechanism underlying the induction of glucose transporter gene expression by the v-src oncogene we studied cell lines that overexpress the normal counterpart of the v-src protein (c-src), or various mutants of the c-src protein. In these mutants, the tyrosines at positions 416, 527, or 519, or various combinations of these, have been replaced by phenylalanine by site directed mutagenesis, resulting in mutated c-src proteins that possess varying tyrosine kinase activity and transforming potential. Cells that overexpress the c-src protein show no changes in glucose transporter gene expression. However, when Tyr 527 in the COOH terminus of the c-src protein is replaced with Phe, the tyrosine kinase activity and transforming potential of the protein are increased and the protein acquires a potent ability to increase levels of glucose transporter mRNA and protein, as well as the rate of 2-deoxy-D-glucose uptake. This ability is abolished by the double mutation of Tyr to Phe in positions 416 and 527, which reduces the tyrosine kinase activity of the 527 single mutant. Thus, the ability of src proteins to induce expression of the glucose transport system is linked to the tyrosine kinase activity of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

[1]  M. Birnbaum.  Identification of a novel gene encoding an insulin-responsive glucose transporter protein , 1989, Cell.

[2]  L. Williams.  Signal transduction by the platelet-derived growth factor receptor. , 1989, Science.

[3]  D. James,et al.  Molecular cloning and characterization of an insulin-regulatable glucose transporter , 1989, Nature.

[4]  J. Newport,et al.  Unraveling of mitotic control mechanisms , 1988, Cell.

[5]  B. Rollins,et al.  Platelet-derived growth factor regulates glucose transporter expression. , 1988, The Journal of biological chemistry.

[6]  G I Bell,et al.  Evidence for a family of human glucose transporter-like proteins. Sequence and gene localization of a protein expressed in fetal skeletal muscle and other tissues. , 1988, The Journal of biological chemistry.

[7]  H. Lodish,et al.  Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney, and β-pancreatic islet cells , 1988, Cell.

[8]  M. Birnbaum,et al.  Growth factors rapidly induce expression of the glucose transporter gene. , 1988, The Journal of biological chemistry.

[9]  K. Gould,et al.  Platelet-derived growth factor induces multisite phosphorylation of pp60c-src and increases its protein-tyrosine kinase activity , 1988, Molecular and cellular biology.

[10]  H. Hanafusa,et al.  Phosphorylation of cellular proteins in Rous sarcoma virus-infected cells: analysis by use of anti-phosphotyrosine antibodies , 1988, Molecular and cellular biology.

[11]  J. Brugge,et al.  Investigation of factors that influence phosphorylation of pp60c-src on tyrosine 527 , 1988, Molecular and cellular biology.

[12]  B. Sefton,et al.  Identification of multiple novel polypeptide substrates of the v-src, v-yes, v-fps, v-ros, and v-erb-B oncogenic tyrosine protein kinases utilizing antisera against phosphotyrosine. , 1988, Oncogene.

[13]  D. Shalloway,et al.  Altered phosphorylation and activation of PP60c-src during fibroblast mitosis , 1988, Cell.

[14]  W. Nelson,et al.  Elevated expression of pp60c-src alters a selective morphogenetic property of epithelial cells in vitro without a mitogenic effect , 1988, Molecular and cellular biology.

[15]  B. Kahn,et al.  Mechanism for markedly hyperresponsive insulin-stimulated glucose transport activity in adipose cells from insulin-treated streptozotocin diabetic rats. Evidence for increased glucose transporter intrinsic activity. , 1987, The Journal of biological chemistry.

[16]  T. Kmiecik,et al.  Activation and suppression of pp60c-src transforming ability by mutation of its primary sites of tyrosine phosphorylation , 1987, Cell.

[17]  C. Cartwright,et al.  Cell transformation by pp60c-src mutated in the carboxy-terminal regulatory domain , 1987, Cell.

[18]  T. Roberts,et al.  Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src , 1987, Cell.

[19]  T. Hunter.  A tail of two src's: Mutatis mutandis , 1987, Cell.

[20]  H. Lodish,et al.  Elevated levels of glucose transport and transporter messenger RNA are induced by ras or src oncogenes. , 1987, Science.

[21]  M. Birnbaum,et al.  Transformation of rat fibroblasts by FSV rapidly increases glucose transporter gene transcription. , 1987, Science.

[22]  M. Birnbaum,et al.  Cloning and characterization of a cDNA encoding the rat brain glucose-transporter protein. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[23]  T. Roberts,et al.  Retrovirus shuttle vector for study of kinase activities of pp60c-src synthesized in vitro and overproduced in vivo , 1986, Molecular and cellular biology.

[24]  Jonathan A. Cooper,et al.  Tyr527 is phosphorylated in pp60c-src: implications for regulation. , 1986, Science.

[25]  A. Markham,et al.  Mutations around the NG59 lesion indicate an active association of polyoma virus middle‐T antigen with pp60c‐src is required for cell transformation. , 1986, The EMBO journal.

[26]  H. Lodish,et al.  Sequence and structure of a human glucose transporter. , 1985, Science.

[27]  S. Alemà,et al.  Differentiation of PC12 phaeochromocytoma cells induced by v-src oncogene , 1985, Nature.

[28]  P. Yaciuk,et al.  Overexpression of the c-src protein does not induce transformation of NIH 3T3 cells. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Parsons,et al.  Monoclonal antibodies to Rous sarcoma virus pp60src react with enzymatically active cellular pp60src of avian and mammalian origin , 1984, Journal of virology.

[30]  E. Gotschlich,et al.  A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. , 1984, Analytical biochemistry.

[31]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[32]  D. Baltimore,et al.  Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus , 1983, Cell.

[33]  H. Hanafusa,et al.  Structure and sequence of the cellular gene homologous to the RSV src gene and the mechanism for generating the transforming virus , 1983, Cell.

[34]  T. Hunter,et al.  Transforming gene product of Rous sarcoma virus phosphorylates tyrosine , 1980, Proceedings of the National Academy of Sciences.

[35]  M Hatanaka,et al.  Transport of sugars in tumor cell membranes. , 1974, Biochimica et biophysica acta.

[36]  G. M. Richards.  Modifications of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA. , 1974, Analytical biochemistry.