Abstract
Although it is well established that cellular transformation with tumor virus leads to changes on glucose metabolism, the effects of cell infection by non-transforming virus are far to be completely elucidated. In this study, we report the first evidence that cultured Vero cells infected with the alphavirus Mayaro show several alterations on glucose metabolism. Infected cells presented a two fold increase on glucose consumption, accompanied by an increment in lactate production. This increase in glycolytic flux was also demonstrated by a significant increase on the activity of 6-phosphofructo 1-kinase, one of the regulatory enzymes of glycolysis. Analysis of the kinetic parameters revealed that the regulation of 6-phosphofructo 1-kinase is altered in infected cells, presenting an increase in V max along with a decrease in K m for fructose-6-phosphate. Another fact contributing to an increase in enzyme activity was the decrease in ATP levels observed in infected cells. Additionally, the levels of fructose 2,6-bisphosphate, a potent activator of this enzyme, was significantly reduced in infected cells. These observations suggest that the increase in PFK activity may be a compensatory cellular response to the viral-induced metabolic alterations that could lead to an impairment of the glycolytic flux and energy production. (Mol Cell Biochem 266: 191–198, 2004)
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Barrett MP, Walmsley AR, Gould GW: Structure and function of facilitative sugar transporters. Curr Opin Cell Biol 11(4): 496–502, 1999
Shepherd PR, Kahn BB: Glucose transporters and insulin action-implications for insulin resistance and diabetes mellitus. New Engl J Med 341(4): 248–257, 1999
Zierler K: Whole body glucose metabolism. Am J Physiol 276 (3 Pt 1): E409–E426, 1999
Hatanaka M, Huebner RJ, Gilden RV: Alterations in the characteristics of sugar uptake by mouse cells transformed by murine sarcoma viruses. J Natl Cancer Inst 43(5): 1091–1096, 1969
Hatanaka M, Augl C, Gilden RV: Evidence for a functional change in the plasma membrane of murine sarcoma virus-infected mouse embryo cells. J Biol Chem 245(4): 714–771, 1970
Weber MJ: Hexose transport in normal and in rous sarcoma virus-transformed cells. J Biol Chem 248(9): 2978–2983, 1973
Hatanaka M, Hanafusa H: Analysis of a functional change in membrane in the process of cell transformation by rous sarcoma virus; alteration in the characteristics of sugar transport. Virology 41(4): 647–652,1970
Birnbaum MJ, Haspel HC, Rosen OM: Transformation of rat fibroblastsby FSV rapidly increases glucose transporter gene transcription.Science 235(4795): 1495–1498, 1987
Young AT, Dahl J, Hausdorff SF, Bauer P H, Birnbaum MJ, Benjamin TL: Phosphatidylinositol 3-kinase binding to polyoma virus middle tumour antigen mediates elevation of glucose transport by increasing translocation of the GLUT1 transporter. Proc Natl Acad Sci USA 92(25):11613–11617, 1995
Singh M, Singh VN, August JT, Horecker BL: Alterations in glucose metabolism in chick embryo cells transformed by rous sarcoma virus. Arch Biochem Biophys 165(1): 240–246, 1974
Singh VN, Singh M, August JT, Horecker BL: Alterations in glucose metabolism in chick-embryo cells transformed by rous sarcoma virus: Intracellular levels of glycolytic intermediates. Proc Natl Acad Sci USA 71(10): 4129–4132, 1974
Gregory SH, Kumari HL, Lakshmi MV, Bose SK: Glycolytic enzymeactivities in murine sarcoma virus-transformed cultures of balb 3T3. Arch Biochem Biophys 175(2): 644–653, 1976
Bosca L, Mojena M, Ghysdael J, Rousseau GG, Hue L: Expression of the v-src or v-fps oncogene increases fructose 2,6-bisphosphate in chick-embryo fibroblasts. Biochem J 236(2): 595–599, 1986
Abreu SL, Lucas-Lenard J: Cellular protein synthesis shutoff by mengovirus: translation of nonviral and viral mRNA's in extracts from un-infected and infected Ehrlich ascites tumour cells. J Virol 18(1): 182–194, 1976
Jen G, Detjen BM, Thach RE: Shutoff of HeLa cell protein synthesis by encephalomyocarditis virus and poliovirus: A comparative study. JVirol 35(1): 150–156, 1980
van Steeg H, Thomas A, Verbeek S, Kasperaitis M, Voorma HO, Benne HO: Shutoff of neuroblastoma cell protein synthesis by Semliki Forest virus: Loss of ability of crude initiation factors to recognize early Semliki Forest virus and host mRNA's. J Virol 38(2): 728–736,1981
Sorbara LR, Maldarelli F, Chamoun G, Schilling B, Chokekijcahi S, Staudt L, Mitsuya H, Simpson IA, Zeichner SL: Human immunodeficiency virus type 1 infection of H9 cells induces in-creased glucose transporter expression. J Virol 70(10): 7275–7279,1996
Gray MA, Micklem KJ, Brown F, Pasternak CA: Effects of vesicular stomatitis virus and semliki forest virus on the uptake of nutrients and intracellular cation concentration. J Gen Virol 64(Pt 7): 1449–1456,1983
Garry RF, Bostick DA, Ulug ET: Sindbis virus infection increases hexose transport in quiescent cells. Virology 155(2): 378–391,1986
Widnell CC, Baldwin AS, Davies A, Martin S, Pasternak CA: Cellular stress induces a redistribution of the glucose transporter. FASEB J 4(6):1634–1637, 1990
Barroso MMS, Lima CS, Silva-Neto MA, Da Poian AT: Mayaro virus infection cycle relies on casein kinase 2 activity. Biochem Biophys Res Commun 296(5): 1334–1339, 2002
Sola-Penna M, dos Santos AC, Alves GG, El-Bacha T, Faber-Barata J, Pereira MF, Serejo FC, Da Poian AT, Sorenson M: A radioassay for phosphofructokinase-1 activity in cell extracts and purified enzyme. J Biochem Biophys Methods 50(2–3): 129–140, 2002
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with folin phenol reagent. J Biol Chem 193: 265–275, 1951
Yang NC, Ho WM, Chen YH, Hu ML: A convenient one-step extraction of cellular ATP using boiling water for the luciferin-luciferase assay of ATP. Anal Biochem 306(2): 323–327, 2002
Van Schaftingen E: Fructose 2,6-bisphosphate. Adv Enzymol Relat Areas Mol Biol 59: 315–395, 1987
Pasternak CA, Whitaker-Dowling PA, Widnell CC: Stress-induced increased of hexose transport as a novel index of cytopathic effects in virus-infected cells: Role of the L protein in the action of vesicular stomatitis virus. Virology 166(2): 379–386, 1988
Lange K, Brandt U, Keller K, Zimmermann B: Endogenous regulation of 2-deoxyglucose uptake in C6 glioma cells correlates with cytoskeleton-mediatedchanges of surface morphology. J Cell Physiol 140(1): 29–43,1989
Quraishi AN, Illsley NP: Transport of sugars across human placental membranes measured by light scattering. Placenta 20(2–3): 167–174,1999
Caro C, Colby-Germinario S, Brenner B, Oliveira M, Wainberg MA, Germinario RJ: Sugar transport and glut transporter expression in a variety of human immunodeficiency virus-1 (HIV-1) chronically infected target cell lines. Int J Biochem Cell Biol 30(9): 1031–1038, 1998
Mazurek S, Zwerschke W, Jansen-Durr P, Eigenbrodt E: Effects of thehuman papilloma virus HPV-16 E7 oncoprotein on glycolysis and glutaminolysis: Role of pyruvate kinase type M2and the glycolytic-enzyme complex. Biochem J 356(Pt1): 247–256, 2001
Romero LM, Raley-Susman KM, Redish DM, Brooke SM, Horner HC, Sapolsky RM: Possible mechanisms by which stress accelerates growth of virally derived tumours. Proc Natl Acad Sci USA 89(22):11084–11087, 1992
Lachmi B, Kaariainen L: Control of protein synthesis in Semliki forest virus-infected cells. J Virol 22(1): 142–149, 1977
Garry RF, Bishop JM, Parker S, Westbrook K, Lewis G, Waite MR: Na+ and K+ concentrations and the regulation of protein synthesis in Sindbis virus-infected chick cells. Virology 96(1): 108–120,1979
Mezencio JMS, Rebello MA: Mayaro virus proteins. MemInst Oswaldo Cruz 88(2): 299–304, 1993
Burlandy FM, Rebello MA: Inhibition of Mayaro virus replication by prostaglandin A(1) in Vero cells. Intervirology 44(6): 344–349,2001
Bardell D, Essex M: Glycolysis during early infection of feline and human cells with feline leukemia virus. Infect Immun 9(5): 824–827,1974
Vora S, Halper JP, Knowles DM: Alterations in the activity and isozymic profile of human PFK during malignant Transformation in vivo and in vitro: Transformation-and progressions-linked discriminants of malignancy. Cancer Research 45(7): 2993–3001,1985
Venuta S, Rubin H: Sugar transport in normal and Rous sarcoma virus-transformedchick-embryo fibroblasts. Proc Natl Acad Sci USA 70(3): 653–657, 1973
Uyeda K: Phosphofructokinase. Adv Enzymol Relat Areas Mol Biol 48: 193–244, 1979
Bardeletti G: Respiration and ATP level in BHK21/13S cells during the earliest stages of rubella virus replication. Intervirology 8(2): 100–109,1977
Terekhina NA, Petrovich I, Parkhomenko TG: Glycolytic metabolites and adenosine triphosphoric acid in the herpes-infected eye. Zh Mikrobiol Epidemiol Immunobiol 2: 92–94, 1998
Pilkis SJ, El-Maghrabi MR, Pilkis J, Claus TH, Cumming DA: Fructose 2,6-bisphosphate: A new activator of phosphofructokinase. J Biol Chem 256(7): 3171–3174, 1981
Gomes-Alves G, Sola-Penna M: Epinephrine modulates cellular distribution of muscle phosphofructokinase. Mol Genet Metab 78(4): 302–306, 2003
El-Bacha T, de Freitas MS, Sola-Penna M: Cellular distribution of phosphofructokinase activity and implications to metabolic regulation in human breast cancer. Mol Genet Metab 79(4): 294–299, 2003
Penso J, Beitner R: Detachment of glycolytic enzymes from cytoskeleton of Lewis lung carcinoma and colon adenocarcinoma cells induced by clotrimazole and its correlation to cell viability and morphology. Mol Genet Metab 76(3): 181–188, 2002
Ovádi J, Orosz F: A new concept for control of glycolysis. In: L. Agiuns, H.S.A. Sherratt (eds.). Channeling in Intermediary Metabolism. Portland Press Research Monograph IX, Portland Press Ltd., London, 1997, pp 237–268
Roberts SJ, Somero GN: Binding of phosphofructokinase to filamentous actin. Biochemistry 26(12): 3437–3442, 1987
Baggetto LG: Deviant energetic metabolism of glycolytic cancer cells. Biochimie 74(11): 959–974, 1992
Sánchez-Martínez C, Aragón JJ: Analysis of phosphofructokinase sub-units and isozymes in ascites tumour cells and its original tissue, murine mammary gland. FEBS Lett. 409(1): 86–90, 1997
Riera M, Roher N, Miro F, Gil C, Trujillo R, Aguilera J, Plana M, Itarte E: Association of protein kinase CK2 with eukaryotic translation initiation factor eIF-2 and with grp94/endoplasmin. Mol Cell Biochem 191(1–2): 97–104, 1999
Meggio F, Pinna LA: One-thousand-and-one substrates of protein kinase CK2? FASEB J 17(3): 349–368, 2003
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El-Bacha, T., Menezes, M.M., Azevedo e Silva, M.C. et al. Mayaro virus infection alters glucose metabolism in cultured cells through activation of the enzyme 6-phosphofructo 1-kinase. Mol Cell Biochem 266, 191–198 (2004). https://doi.org/10.1023/B:MCBI.0000049154.17866.00
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DOI: https://doi.org/10.1023/B:MCBI.0000049154.17866.00