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Different regulation of mitochondrial apoptosis and bcl-2 gene expression in quiescent and proliferative human fibroblasts infected with cytomegalovirus

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Abstract

The goal of this work was to compare the dynamics of mitochondrial apoptosis (MA) in cells at different stages of proliferation and with different susceptibilities to cytomegalovirus (CMV). It was found that, in quiescent human fibroblasts (HFs), this virus can regulate MA at the level of bcl-2 gene transcription in two directions by causing both pro- and anti-apoptotic effects. The suppression of bcl-2 transcription is stronger in the HF-977 line, which is highly susceptible to CMV, than in the HF-1608 line. The effect of proliferative activity on MA was studied on the CMV-infected HF-110044 line at the G0 or S phase. A direct correlation was established between the accumulation of cytochrome c and caspase-3 (MA markers) and the production of IE72, pp65, and gB (CMV proteins). In G0-fibroblasts, viral replication was highly productive and the bcl-2 expression was ten times higher than in the S-phase cells whose viral protein production and cell death were much lower. An increase of gene transcription and the accumulation of Bcl-2 protein enhanced cell viability and enabled the synthesis of viral proteins. The impairment of the structure of actin microfilaments, a target of caspase-3, coincided with the pronounced suppression of the gamma-actin gene in the Sphase HF-110044. Our findings unravel the CMV-induced MA mechanisms that lead to the rapid death of infected quiescent fibroblasts and slow down the death of cells infected at the stage of DNA synthesis.

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Abbreviations

RT:

reverse transcriptase

RT-PCR:

polymerase chain reaction (PCR) using cDNA synthesized by RT as a matrix

CMV:

human cytomegalovirus

CMVI:

cytomegaloviral infection

PB:

phosphate buffer

HF:

human fibroblasts

FCS:

fetal calf serum

References

  • Andoniou, C.E. and Digli-Esposti, M.A., Insights into the Mechanisms of Cmv-Mediated Interference with Cellular Apoptosis, Immunol. Cell Biol., 2006, vol. 84, pp. 99–106.

    Article  CAS  PubMed  Google Scholar 

  • Arnoult, D., Bartle, L.M., Skaletskaya, A., Porcet, D., Zamzami, N., Park, P.U., Sharpe, J., Youle, R.J., and Goldmacher, V.S., Cytomegalovirus Cell Death Suppressor VMIA Blocks Bax but not Bak Mediated Apoptosis by Binding and Sequestering Bax at Mitochondira, Proc. Natl. Acad. Sci. USA, 2004, vol. 101, pp. 7988–7993.

    Article  CAS  PubMed  Google Scholar 

  • Belkhiri, A., Dar, A.A., Zaika, A., Kelly, M., and El-Rifai, W., Tdarpp Promotes Cancer Cell Survival by up-Regulation of Bc12 through Akt-Dependent Mechanism, Cancer Res., 2008, vol. 68, pp. 395–403.

    Article  CAS  PubMed  Google Scholar 

  • Bigelow, R.L.H., Chari, N.S., Unden, A.B., Spurgers, K.B., Lee, S., Roop, D.R., Toftgard, R., and Mcdonnell, T.J., Transcriptional Regulation of Bcl-2 Mediated by the Sonic Hedgehog Signaling Pathway through Gli-1, J. Biol. Chem., 2004, vol. 279, pp. 1197–1205.

    Article  CAS  PubMed  Google Scholar 

  • Bystrevskaya, V.B., Lobova, T.V., Smirnov, V.N., Makarova, N.E., and Kushch, A.A., Centrosome Injury in Cells Infected with Human Cytomegalovirus, J. Struct. Biol., 1997, vol. 120, pp. 52–60.

    Article  CAS  PubMed  Google Scholar 

  • Casavant, N.C., Luo, M.H., Rosenke, K., Winegardner, T., Zurawska, A., and Fortunato, E.A., Potential Role for P53 in the Permissive Life Cycle of Human Cytomegalovirus, J. Virol., 2006, vol. 80, pp. 8390–8401.

    Article  CAS  PubMed  Google Scholar 

  • Castillo, J.P., and Kowalik, T.F., Human Cytomegalovirus Immediate Early Proteins and Cell Growth Control, Gene, 2002, vol. 290, pp. 19–34.

    Article  CAS  PubMed  Google Scholar 

  • Castillo, J.P., Frame, F.M., Rogoff, H.A., Pickering, M.T., Yurochko, A.D., and Kowalik, T.F., Human Cytomegalovirus IE1-72 Activates Ataxia Telangiectasia Mutated Kinase and a P53/P21-Mediated Growth Arrest Response, J. Virol., 2005, vol. 79, pp. 11467–11475.

    Article  CAS  PubMed  Google Scholar 

  • Castillo, J.P., Yurochko, A.D., and Kowallik, T.F., Role of Human Cytomegalovirus Immediate-Early Proteins in Cell Growth Control, J. Virol., 2000, vol. 74, pp. 8028–8037.

    Article  CAS  PubMed  Google Scholar 

  • Challacombe, J.F., Rechtsteiner, A., Gottardo, R., Rocha, L.M., Browne, E.P., Shenk, T., Alther, M.R., and Brettin, T.S., Evalution of the Host Transcriptional Response to Human Cytomegalovirus Infection, Physiol. Genomics, 2004, vol. 18, pp. 5–62.

    Google Scholar 

  • Chipuk, J.E. and Green, D.R., How Do Bcl-2 Proteins induce Mitochondrial Outer Membrane Permeabilization, Trends Cell Biol., 2008, vol. 18, pp. 157–164.

    Article  CAS  PubMed  Google Scholar 

  • Chomczynski, P. and Sacchi, N., Single-Step Method of RNA Isolation by Acid Guanidinium Thiocyanate-Phenol-Chloroform Extraction, Anal. Biochem., 1987, vol. 162, pp. 156–159.

    Article  CAS  PubMed  Google Scholar 

  • Domnina, L.V., Ivanova, O.Yu., Chernyak, B.V., Skulachev, V.P., and Vasilev, Yu.M., Effects of Inhibitors of Dynamics of Cytoskeletal Structures on the Development of Apoptosis Induced by the Tumor Necrosis Factor, Biokhimiya, 2002, vol. 67, no. 7, pp. 890–900.

    Google Scholar 

  • Erba, H.P., Eddy, R., Shows, T., Kedes, L., and Gunning, P., Structure, Chromosome Location, and Expression of the Human Gamma-Actin Gene: Differential Evolution, Location, and Expression of the Cytoskeletal Beta- and Gamma-Actin Genes, Mol, Cell Biol., 1988, vol. 8, pp. 1775–1789.

    CAS  Google Scholar 

  • Fedorova, N.E., Medzhidova, A.A., Medzhidova, M.G., and Kusch, A.A., Blockade of Cell Proliferation and Mitosis Pathology in Cells Infected by Cytomegalovirus: The Role of the Cell Cycle Period at the Moment of Infection, Dokl. Akademii Nauk, 2003, vol. 392, pp. 471–474.

    CAS  Google Scholar 

  • Fedorova, N.E., Sokolova, T.M., Medzhidova, M.G., Uryvaev, L.V., and Kusch, A.A., CMV-Induced Cell Death and Expression of the Fas Gene in Resting and Proliferating Human Fibroblasts, Zh. Microbiol. (Moscow), 2008, vol. 5, pp. 42–48.

    Google Scholar 

  • Fortunato, E.A., Sanchez, V., Yen, J.Y., and Spector, D.H., Infection of Cells with Human Cytomegalovirus during S Phase Results in a Blockade to Immediate-Early Gene Expression That Can Be Overcome by Inhibition of the Proteasome, J. Virol., 2002, vol. 76, pp. 5369–5379.

    Article  CAS  PubMed  Google Scholar 

  • Ginsberg, D., E2F1 Pathways to Apoptosis, FEBS Letters, 2002, vol. 529, pp. 122–125.

    Article  CAS  PubMed  Google Scholar 

  • Glazunova, V.A. and Shtil, A.A., Mitochondrial Mechanisms of Apoptosis in Response to DNA Damage, Mol. Biol. (Mosc.), 2008, vol. 42, no. 5, pp. 765–771.

    CAS  Google Scholar 

  • Goldmacher, V.S., Cell Death Suppression by Cytomegalovirus, Apoptosis, 2005, vol. 10, pp. 251–265.

    Article  CAS  PubMed  Google Scholar 

  • Gulbins, E., Dreschers, S., and Bock, J., Role of Mitochondria in Apoptosis, Exp. Physiol., 2003, vol. 88, pp. 85–90.

    Article  CAS  PubMed  Google Scholar 

  • Hengartner, M.O., The Biochemistry of Apoptosis, Nature, 2000, vol. 407, pp. 770–776.

    Article  CAS  PubMed  Google Scholar 

  • Hertel, L. and Mocarski, E.S., Global Analysis of Host Cell Gene Expression Late during Cytomegalovirus Infection Reveals Extensive Dysregulation of Cell Cycle Gene Expression and Induction of Pseudomitosis Independent of Us28 Function, J. Virol., 2004, vol. 78, pp. 11 988–12 011.

    Article  CAS  Google Scholar 

  • Johnson, R.A., Wang, X., Ma, X.L., Huang, S.M., and Huang E.S., Human Cytomegalovirus up-Regulates the Phosphatidylinositol 3-Kinase (Pi3-K) Pathway: Inhibition of Pi3-K Activity Inhibits Viral Replication and Virus-Induced Signaling, J. Virol., 2001, vol. 75, pp. 6022–6032.

    Article  CAS  PubMed  Google Scholar 

  • Lin, M.F., Wei, G.Q., Huang, H., and Cai, Z., Human Cytomegalovirus Induces Alteration of Beta-Actin mRNA and Microfilaments in Human Embryo Fibroblast Cells, J. Zhejiang Univ. Sci., 2004, vol. 5, pp. 733–737.

    Article  CAS  PubMed  Google Scholar 

  • Luo, H.R., Hattori, H., Hossain, M.A., Hester, L., Huang, Y., Lee-Kwon, W., Donowitz, M., Nagata, E., and Snyder, S.H., Akt As a Mediator of Cell Death, Proc. Natl. Acad. Sci. USA, 2003, vol. 100, pp. 11712–11717.

    Article  CAS  PubMed  Google Scholar 

  • McCormick, A.L., Control of Apoptosis by Human Cytomegalovirus, Curr. Top. Microbiol. Immunol., 2008, vol. 325, pp. 281–295.

    Article  CAS  PubMed  Google Scholar 

  • Paulus, C., Krauss, S., and Nevels, M., A Human Cytomegalovirus Antagonist of Type I Ifn-Dependent Signal Transducer and Activator of Transcription Signaling, Proc. Natl. Acad. Sci. USA, 2006, vol. 103, pp. 3840–3845.

    Article  CAS  PubMed  Google Scholar 

  • Reboredo, M., Greaves, R.F., and Hahn, G., Human Cytomegalovirus Proteins Encoded by Ul37 Exon 1 Protect Infected Fibroblasts Against Virus-Induced Apoptosis and Are Required for Efficient Virus Replication, J. Gen. Virol., 2004, vol. 85, pp. 3555–3567.

    Article  CAS  PubMed  Google Scholar 

  • Schroeder, B.M., Christensen, R., and Worthen, G.S., Human Cytomegalovirus Protects Endothelial Cells from Apoptosis Induced by Growth Factor Withdrawal, J. Clin. Virol., 2002, vol. 25,Suppl. 2, pp. S149–S157.

    Google Scholar 

  • Sciortino, M.T., Perry, D., Medici, M.A., Grelli, S., Serafino, A., Borner, C., and Mastino, A., Role of Bcl-2 Expression for Productive Herpes Simplex Virus 2 Replication, Virology, 2006, vol. 356, pp. 136–146.

    Article  CAS  PubMed  Google Scholar 

  • Skaletskaya, A., Bartle, L.M., Chitenden, T., McCormick, A.I., Mocarski, E.S., and Goldmacher, V.S., A Cytomegalovirus-Encoded Inhibitor of Apoptosis That Suppresses Caspase-8 Activation, Proc. Natl. Acad. Sci. USA, 2001, vol. 98, pp. 7828–7834.

    Article  Google Scholar 

  • Sokolova, T.M., Bibikova, O.V., Bystrov, N.S., and Uryvaev, L.V., Expression of Genes of the Interferon System and Cell Apoptosis in Human Blood Samples, Vopr. Virusol., 2005, vol. 50, no. 1, pp. 19–23.

    CAS  PubMed  Google Scholar 

  • Sokolova, T.M., Fedorova, N.E., Medzhidova, M.G., Terekhov, S.M., Uryvaev, L.V., and Kusch, A.A., Regulation of the Level of 2′,5′-Oligoadenylate Synthetase Gene Activity by Cytomegalovirus in Human Fibroblasts, Vopr. Virusol., 2007a, vol. 52, vol. 1, pp. 28–36.

    CAS  PubMed  Google Scholar 

  • Sokolova, T.M., Fedorova, N.E., Medzhidova, M.G., Terekhov, S.N., Uryvaev, L.V., and Kusch, A.A., Mechanisms of Cell Resistance to Cytomegalovirus Are Related to the Transcriptional Activity of Genes of Leukocytic and Immune Interferons, Med. Immunol., 2007b, vols. 4–5, pp. 457–466.

    Google Scholar 

  • Tsujimoto, Y. and Croce, C.M., Analysis of the Structure, Transcripts, and Protein Products of Bcl-2, the Gene Involved in Human Follicular Lymphoma, Proc. Natl. Acad. Sci. USA, 1986, vol. 83, pp. 5214–5218.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, Y., Weyman, C.M., Liu, H., Almasan, A., and Zhou, A., Ifn-Gamma Induces Apoptosis in Hl-60 Cells through Decreased Bcl-2 and Increased Bak Expression, J. Interferon Cytokine Res., 2008, vol. 28, pp. 65–72.

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Ivanovskii Institute of Virology, Russian Academy of Medical Sciences, Moscow, Russia

    N. E. Fedorova, T. M. Sokolova, M. G. Medzhidova & A. A. Kushch

Authors
  1. N. E. Fedorova
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  2. T. M. Sokolova
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  3. M. G. Medzhidova
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  4. A. A. Kushch
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Corresponding author

Correspondence to T. M. Sokolova.

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Original Russian Text © N.E. Fedorova, T.M. Sokolova, M.G. Medzhidova, A.A. Kushch, 2010, published in Tsitologiya, Vol. 52, No. 2, 2010, pp. 168–176.

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Fedorova, N.E., Sokolova, T.M., Medzhidova, M.G. et al. Different regulation of mitochondrial apoptosis and bcl-2 gene expression in quiescent and proliferative human fibroblasts infected with cytomegalovirus. Cell Tiss. Biol. 4, 143–151 (2010). https://doi.org/10.1134/S1990519X10020045

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  • DOI: https://doi.org/10.1134/S1990519X10020045

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