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. 1992 Jul 15;285(Pt 2):413–417. doi: 10.1042/bj2850413

Activation of 6-phosphofructo-2-kinase by pp60v-src is an indirect effect.

M J Marchand 1, L Maisin 1, L Hue 1, G G Rousseau 1
PMCID: PMC1132804  PMID: 1322131

Abstract

6-Phosphofructo-2-kinase (PFK-2) catalyses the synthesis of fructose 2,6-bisphosphate (Fru-2,6-P2), a potent stimulator of glycolysis. In chick-embryo fibroblasts, PFK-2 activity and Fru-2,6-P2 concentration increase upon transformation by Rous sarcoma virus. We show here that the increase in PFK-2 activity required more than 2 h after shifting fibroblasts infected with a thermosensitive mutant of Rous sarcoma virus from the restrictive to the permissive temperature. Pretreatment of the cells with actinomycin D prevented this increase in PFK-2 activity, suggesting a requirement for RNA synthesis. However, the increase in PFK-2 activity did not correspond to an increase in immunoprecipitable PFK-2. Moreover, the thermostability of PFK-2 and the affinity of this enzyme for its substrate fructose 6-phosphate were increased upon transformation by Rous sarcoma virus. Staurosporine, an inhibitor of protein kinase C, prevented the increase in PFK-2 activity brought about by the shift to the permissive temperature. This, together with a comparison of the effects of phorbol esters on PFK-2 activity, suggests that pp60v-src stimulates, via protein kinase C, the transcription of a gene whose products is a distinct PFK-2 isoenzyme or a protein that activates PFK-2.

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Selected References

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  1. Bartrons R., Hue L., Van Schaftingen E., Hers H. G. Hormonal control of fructose 2,6-bisphosphate concentration in isolated rat hepatocytes. Biochem J. 1983 Sep 15;214(3):829–837. doi: 10.1042/bj2140829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bosca L., Mojena M., Ghysdael J., Rousseau G. G., Hue L. Expression of the v-src or v-fps oncogene increases fructose 2,6-bisphosphate in chick-embryo fibroblasts. Novel mechanism for the stimulation of glycolysis by retroviruses. Biochem J. 1986 Jun 1;236(2):595–599. doi: 10.1042/bj2360595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bosca L., Rousseau G. G., Hue L. Phorbol 12-myristate 13-acetate and insulin increase the concentration of fructose 2,6-bisphosphate and stimulate glycolysis in chicken embryo fibroblasts. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6440–6444. doi: 10.1073/pnas.82.19.6440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Cooper J. A., Reiss N. A., Schwartz R. J., Hunter T. Three glycolytic enzymes are phosphorylated at tyrosine in cells transformed by Rous sarcoma virus. Nature. 1983 Mar 17;302(5905):218–223. doi: 10.1038/302218a0. [DOI] [PubMed] [Google Scholar]
  6. Crepin K. M., Darville M. I., Hue L., Rousseau G. G. Characterization of distinct mRNAs coding for putative isozymes of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Eur J Biochem. 1989 Aug 1;183(2):433–440. doi: 10.1111/j.1432-1033.1989.tb14946.x. [DOI] [PubMed] [Google Scholar]
  7. Darville M. I., Chikri M., Lebeau E., Hue L., Rousseau G. G. A rat gene encoding heart 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. FEBS Lett. 1991 Aug 19;288(1-2):91–94. doi: 10.1016/0014-5793(91)81009-w. [DOI] [PubMed] [Google Scholar]
  8. Darville M. I., Crepin K. M., Hue L., Rousseau G. G. 5' flanking sequence and structure of a gene encoding rat 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6543–6547. doi: 10.1073/pnas.86.17.6543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Diringer H., Friis R. R. Changes in phosphatidylinositol metabolism correlated to growth state of normal and Rous sarcoma virus-transformed Japanese quail cells. Cancer Res. 1977 Sep;37(9):2979–2984. [PubMed] [Google Scholar]
  10. Friis R. R. Temperature-sensitive mutants of avian RNA tumor viruses: a review. Curr Top Microbiol Immunol. 1978;79:261–293. doi: 10.1007/978-3-642-66853-1_6. [DOI] [PubMed] [Google Scholar]
  11. Hue L., Rider M. H. Role of fructose 2,6-bisphosphate in the control of glycolysis in mammalian tissues. Biochem J. 1987 Jul 15;245(2):313–324. doi: 10.1042/bj2450313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson R. M., Wasilenko W. J., Mattingly R. R., Weber M. J., Garrison J. C. Fibroblasts transformed with v-src show enhanced formation of an inositol tetrakisphosphate. Science. 1989 Oct 6;246(4926):121–124. doi: 10.1126/science.2506643. [DOI] [PubMed] [Google Scholar]
  13. Jove R., Hanafusa H. Cell transformation by the viral src oncogene. Annu Rev Cell Biol. 1987;3:31–56. doi: 10.1146/annurev.cb.03.110187.000335. [DOI] [PubMed] [Google Scholar]
  14. Kawai S., Hanafusa H. The effects of reciprocal changes in temperature on the transformed state of cells infected with a rous sarcoma virus mutant. Virology. 1971 Nov;46(2):470–479. doi: 10.1016/0042-6822(71)90047-x. [DOI] [PubMed] [Google Scholar]
  15. Martins T. J., Sugimoto Y., Erikson R. L. Dissociation of inositol trisphosphate from diacylglycerol production in Rous sarcoma virus-transformed fibroblasts. J Cell Biol. 1989 Feb;108(2):683–691. doi: 10.1083/jcb.108.2.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mojena M., Bosca L., Hue L. Effect of glutamine on fructose 2,6-bisphosphate and on glucose metabolism in HeLa cells and in chick-embryo fibroblasts. Biochem J. 1985 Dec 1;232(2):521–527. doi: 10.1042/bj2320521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Presek P., Reinacher M., Eigenbrodt E. Pyruvate kinase type M2 is phosphorylated at tyrosine residues in cells transformed by Rous sarcoma virus. FEBS Lett. 1988 Dec 19;242(1):194–198. doi: 10.1016/0014-5793(88)81014-7. [DOI] [PubMed] [Google Scholar]
  18. Rider M. H., Vandamme J., Lebeau E., Vertommen D., Vidal H., Rousseau G. G., Vandekerckhove J., Hue L. The two forms of bovine heart 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase result from alternative splicing. Biochem J. 1992 Jul 15;285(Pt 2):405–411. doi: 10.1042/bj2850405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Singh M., Singh V. N., August J. T., Horecker B. L. Alterations in glucose metabolism in chick embryo cells transformed by Rous sarcoma virus. Transformation-specific changes in the activities of key enzymes of the glycolytic and hexose monophosphate shunt pathways. Arch Biochem Biophys. 1974 Nov;165(1):240–246. doi: 10.1016/0003-9861(74)90160-x. [DOI] [PubMed] [Google Scholar]
  20. Spangler R., Joseph C., Qureshi S. A., Berg K. L., Foster D. A. Evidence that v-src and v-fps gene products use a protein kinase C-mediated pathway to induce expression of a transformation-related gene. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7017–7021. doi: 10.1073/pnas.86.18.7017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tamaoki T., Nomoto H., Takahashi I., Kato Y., Morimoto M., Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. doi: 10.1016/0006-291x(86)90008-2. [DOI] [PubMed] [Google Scholar]
  22. Taylor D. J., Evanson J. M., Woolley D. E. Contrasting effects of the protein kinase C inhibitor, staurosporine, on cytokine and phorbol ester stimulation of fructose 2,6-bisphosphate and prostaglandin E production by fibroblasts in vitro. Comparative studies using interleukin-1 alpha, tumour necrosis factor alpha, transforming growth factor beta, interferon-gamma and 12-O-tetradecanoylphorbol 13-acetate. Biochem J. 1990 Aug 1;269(3):573–577. doi: 10.1042/bj2690573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Van Schaftingen E., Hers H. G. Purification and properties of phosphofructokinase 2/fructose 2,6-bisphosphatase from chicken liver and from pigeon muscle. Eur J Biochem. 1986 Sep 1;159(2):359–365. doi: 10.1111/j.1432-1033.1986.tb09876.x. [DOI] [PubMed] [Google Scholar]
  24. Weinhouse S. The Warburg hypothesis fifty years later. Z Krebsforsch Klin Onkol Cancer Res Clin Oncol. 1976;87(2):115–126. doi: 10.1007/BF00284370. [DOI] [PubMed] [Google Scholar]
  25. Yamada K., Tillotson L. G., Isselbacher K. J. Regulation of hexose carriers in chicken embryo fibroblasts. Effect of glucose starvation and role of protein synthesis. J Biol Chem. 1983 Aug 25;258(16):9786–9792. [PubMed] [Google Scholar]

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