Biopolym. Cell. 2010; 26(3):194-199.
Structure and Function of Biopolymers
Relations between nitric oxide synthase DNOS1, Hsp70 and apoptosis regulatory gene grim in Drosophila melanogaster after heat stress induction
1Khussainova E. M., 1Bulentayeva Z. A., 1Bekmanov B. O., 1Djansugurova L. B., 1Bersimbayev R. I.
  1. Institute of General Genetics and Cytology
    75A, al-Farabi Ave., Almaty, Kazakhstan, 050060

Abstract

Aim. To investigate correlation between nitric oxide synthase, heat shock protein Hsp70 and apoptosis regulatory gene grim in D. melanogaster. Methods. The heat stress (37 C for 1 hour) induction in third instar larvae of Oregon R strain and transgenic strains, containing additional copies of dNOS1 gene. RT-PCR and Western-blot analysis were used to study the expression of dNOS, Hsp70 and grim genes. Results. It is demonstrated that additional copies of dNOS1 gene in transgenic strains are intensively expressed immediately after heat stress induction. It was revealed that in all Drosophila strains the level of Hsp70 gene expression and its protein synthesis increase with subsequent decline after 2–3 hours; whereas the level of expression of grim increases immediately after heat stress induction in transgenic strains and declines in wild type flies, while the level of Hsp70 expression remains high. Conclusions. The increased level of Hsp70 has negative impact on the expression of grim, whereas additional NO synthesis neutralizes anti-apoptotic effects of Hsp70 and increases the expression level of grim. Thus, we assume the competitive relationships between anti-apoptotic functions of Hsp70 and pro-apoptotic effects of nitric oxide.
Keywords: apoptosis, Drosophila, heat shock proteins, nitric oxide, grim

References

[1] Richardson H., Kumar S. Death to flies: Drosophila as a model system to study programmed cell death J. Immunol. Meth 2002 265, N 1–2:21–38.
[2] Kornbluth S., White K. Apoptosis in Drosophila: neither fish nor fowl (nor man, nor worm) J. Cell Sci 2005 118, Pt 9 :1779–1787.
[3] Elmore S. Apoptosis: A review of programmed cell death Toxicol. Pathol 2007 35, N 4:495–516.
[4] Villalobo A. Nitric oxide and cell proliferation FEBS J 2006 273, N 11:2329–2344.
[5] Stasiv Y., Kuzin B., Regulski M., Tully T., Enikolopov G. Regulation of multimers via truncated isoforms: a novel mechanism to control nitric-oxide signaling Genes Develop 2004 18, N 15:1812–1823.
[6] Kim P. K., Zamora R., Petrosko P., Billiar T. R. The regulatory role of nitric oxide in apoptosis Int. Immunopharmacol 2001 1, N 8:1421–1441.
[7] Sreedhar A. S., Csermely P. Heat shock proteins in the regulation of apoptosis: new strategies in tumor therapy: a comprehensive review Pharmacol. Ther 2004 101, N 3 P. 227–257.
[8] Beere H. M. Death versus survival: functional interaction between the apoptotic and stress-inducible heat shock protein pathways J. Clin. Invest 2005 115, N 10:2633–2639.
[9] Arya R., Mallik M., Lakhotia S. C. Heat shock genes – integrating cell survival and death J. Biosci 2007 32, N 3 P. 595–610.
[10] Khlebodarova T. M. How cells protect themselves against stress? Genetika 2002 38, N 4:437–452
[11] Mayer M. P., Bukau B. Hsp70 chaperones: cellular functions and molecular mechanism Cell Mol. Life Sci 2005 62, N 6:670–684.
[12] Song Z., Guan B., Bergman A., Nicholson D. W., Thornberry N. A., Peterson E. P., Steller H. Biochemical and genetic interactions between Drosophila caspases and the proapoptotic genes rpr, hid, and grim Mol. Cell. Biol 2000 20, N 8 P. 2907–2914.
[13] Bangs P., Franc N., White K. Molecular mechanisms of cell death and phagocytosis in Drosophila Cell Death Differ 2000 7, N 11:1027–1034.
[14] Stasiv Y., Regulski M., Kuzin B., Tully T., Enikolopov G. The Drosophila nitric-oxide synthase gene (dNOS) encodes a family of proteins that can modulate NOS activity by acting as dominant negative regulators J. Biol. Chem 2001 276, N 45:42241–42251.
[15] Schmitt E., Gehrmann M., Brunet M., Multhoff G., Garrido C. Intracellular and extracellular functions of heat shock proteins: repercussions in cancer therapy J. Leukoc. Biol 2007 81, N 1:15–27.
[16] Lakhotia S. C., Srivastava P., Prasanth K. V. Regulation of heat shock proteins, Hsp70 and Hsp64, in heat-shocked Malpighian tubules of Drosophila melanogaster larvae Cell Stress Chaperones 2002 7, N 4:347–356.
[17] Krebs R. A., Feder M. E. Tissue specific variation in Hsp70 expression and thermal damage in Drosophila melanogaster larvae J. Exp. Biol. 1997; 200, pt 14:2007–2015.
[18] Guzhova I. V., Arnholdt A. C. V., Darieva Z. A., Kinev A. V., Lasunskaia E. B., Nilsson K., Bozhkova V. M., Voronin A. P., Margulis B. A. The pleiotropic of an extracellular HSP-70 on functional properties of human premonocytes through binding to cell surface and internalization Cell Stress Chaperones 1998 3, N 1:67–77.
[19] McCarthy J. V., Dixit V. M. Apoptosis induced by Drosophila reaper and grim in a human system. Attenuation by inhibitor of apoptosis proteins (cIAPs) J. Biol. Chem 1998 273, N 37:24009–24015.
[20] Tolebaeva A. D., Amirgalieva A. S., Mit N. V., Djansugurova L. B., Bekmanov B.O., Bersimbayev R.I. Development of NO mediated apoptosis regulation model system using oncovirus induced Lobe-mutants of Drosophila melanogaster. Fundamental studies in biology and medicine: collection of scientific papers Stavropol, 2009:1603.