Overexpression of interferon-γ-inducible GTPase inhibits coxsackievirus B3-induced apoptosis through the activation of the phosphatidylinositol 3-kinase/Akt pathway and inhibition of viral replication

Huifang M. Zhang, Ji Yuan, Paul Cheung, Honglin Luo, Bobby Yanagawa, David Chau, Najwan Stephan-Tozy, Brian W. Wong, Jingchun Zhang, Janet E. Wilson, Bruce M. McManus, Decheng Yang

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Abstract

Our previous studies using differential mRNA display have shown that interferon-γ-inducible GTPase (IGTP), was up-regulated in coxsackievirus B3 (CVB3)-infected mouse hearts. In order to explore the effect of IGTP expression on CVB3-induced pathogenesis, we have established a doxycycline-inducible Tet-On HeLa cell line overexpressing IGTP and have analyzed activation of several signaling molecules that are involved in cell survival and death pathways. We found that following IGTP overexpression, protein kinase B/Akt was strongly activated through phosphorylation, which leads to phosphorylation of glycogen synthase kinase-3 (GSK-3). Furthermore, in the presence of CVB3 infection, the intensity of the phosphorylation of Akt was further enhanced and associated with a delayed activation of caspase-9 and caspase-3. These data indicate that IGTP expression appears to confer cell survival in CVB3-infected cells, which was confirmed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl) -2H-tetrazolium salt cell viability assay. However, the ability of IGTP to induce phosphorylation of Akt and to promote cell survival was attenuated by the phosphotidylinositol-3 kinase (PI3-K) inhibitor LY294002. Transient transfection of the cells with a dominant negative Akt construct followed by doxycycline induction and CVB3 infection reversed Akt phosphorylation to basal levels and returned caspase-3 activity to levels similar to those when the PI3-K inhibitor LY294002 was added. Moreover, IGTP expression inhibited viral replication and delayed CVB3-induced cleavage of eukaryotic translation initiation factor 4G, indicating that IGTP-mediated cell survival relies on not only the activation of PI3-K/Akt, inactivation of GSK-3 and suppression of caspase-9 and caspase-3 but also the inhibition of viral replication.

Original languageEnglish (US)
Pages (from-to)33011-33019
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number35
DOIs
StatePublished - Aug 29 2003

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Phosphatidylinositol 3-Kinase
Enterovirus
GTP Phosphohydrolases
Interferons
Phosphorylation
Chemical activation
Apoptosis
Cells
Cell Survival
Coxsackievirus Infections
Caspase 3
Glycogen Synthase Kinase 3
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Phosphotransferases
Caspase 9
Doxycycline
Eukaryotic Initiation Factor-4G
Tetrazolium Salts
Proto-Oncogene Proteins c-akt
Gene Expression Profiling

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Overexpression of interferon-γ-inducible GTPase inhibits coxsackievirus B3-induced apoptosis through the activation of the phosphatidylinositol 3-kinase/Akt pathway and inhibition of viral replication. / Zhang, Huifang M.; Yuan, Ji; Cheung, Paul; Luo, Honglin; Yanagawa, Bobby; Chau, David; Stephan-Tozy, Najwan; Wong, Brian W.; Zhang, Jingchun; Wilson, Janet E.; McManus, Bruce M.; Yang, Decheng.

In: Journal of Biological Chemistry, Vol. 278, No. 35, 29.08.2003, p. 33011-33019.

Research output: Contribution to journalArticle

Zhang, Huifang M. ; Yuan, Ji ; Cheung, Paul ; Luo, Honglin ; Yanagawa, Bobby ; Chau, David ; Stephan-Tozy, Najwan ; Wong, Brian W. ; Zhang, Jingchun ; Wilson, Janet E. ; McManus, Bruce M. ; Yang, Decheng. / Overexpression of interferon-γ-inducible GTPase inhibits coxsackievirus B3-induced apoptosis through the activation of the phosphatidylinositol 3-kinase/Akt pathway and inhibition of viral replication. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 35. pp. 33011-33019.
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abstract = "Our previous studies using differential mRNA display have shown that interferon-γ-inducible GTPase (IGTP), was up-regulated in coxsackievirus B3 (CVB3)-infected mouse hearts. In order to explore the effect of IGTP expression on CVB3-induced pathogenesis, we have established a doxycycline-inducible Tet-On HeLa cell line overexpressing IGTP and have analyzed activation of several signaling molecules that are involved in cell survival and death pathways. We found that following IGTP overexpression, protein kinase B/Akt was strongly activated through phosphorylation, which leads to phosphorylation of glycogen synthase kinase-3 (GSK-3). Furthermore, in the presence of CVB3 infection, the intensity of the phosphorylation of Akt was further enhanced and associated with a delayed activation of caspase-9 and caspase-3. These data indicate that IGTP expression appears to confer cell survival in CVB3-infected cells, which was confirmed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl) -2H-tetrazolium salt cell viability assay. However, the ability of IGTP to induce phosphorylation of Akt and to promote cell survival was attenuated by the phosphotidylinositol-3 kinase (PI3-K) inhibitor LY294002. Transient transfection of the cells with a dominant negative Akt construct followed by doxycycline induction and CVB3 infection reversed Akt phosphorylation to basal levels and returned caspase-3 activity to levels similar to those when the PI3-K inhibitor LY294002 was added. Moreover, IGTP expression inhibited viral replication and delayed CVB3-induced cleavage of eukaryotic translation initiation factor 4G, indicating that IGTP-mediated cell survival relies on not only the activation of PI3-K/Akt, inactivation of GSK-3 and suppression of caspase-9 and caspase-3 but also the inhibition of viral replication.",
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AU - Yuan, Ji

AU - Cheung, Paul

AU - Luo, Honglin

AU - Yanagawa, Bobby

AU - Chau, David

AU - Stephan-Tozy, Najwan

AU - Wong, Brian W.

AU - Zhang, Jingchun

AU - Wilson, Janet E.

AU - McManus, Bruce M.

AU - Yang, Decheng

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