Because viral diseases are major health risks to the society, it is important to understand how they are caused and how our natural defense systems can protect us. A virus-infected cell can either die or it can live;if it lives it may be permanently infected with the virus. This proposal is to investigate the molecular mechanism that dictates the above choice, namely, whether a virus-infected cell dies or lives and produces more viruses. Many major viral diseases are caused by Paramyxoviridae, such as respiratory syncytial virus, parainfluzenza virus, measles virus and mumps virus. The nature and the severity of pathogenesis is determined by the host response, a major component of which is the innate response of the infected cell which may undergo apoptosis or live and be capable to establish persistent infection, the two outcomes having opposite effects on the host organism. We have made the novel observation that in cells not expressing the cellular transcription factor IRF-3, paramyxoviruses are not lytic because IRF-3 initiates an apoptotic response in infected cells. Moreover, this response is greatly accelerated by inhibiting the activity of PI3 kinases. These observations led us to formulate the hypothesis that IRF-3 determines the choice between lytic and non-lytic infection and PI3 kinases determine the timing of the lysis.
Three specific aims will be pursued to test the hypothesis. In the first aim, we will determine which known properties of IRF-3 are needed and which known IRF-3 regulated genes may be mediating apoptosis. These experiments will use over- expression of individual proteins or their ablation by siRNA. Finally, unbiased genetic screens of the whole human genome will be conducted by using insertional mutagenesis or an appropriate shRNA library and cell survival as the end point assay. In the second aim, we will determine whether the intrinsic or the extrinsic pathway is used in viral apoptosis. We will also identify the specific isozyme of PI3 kinase and its downstream targets that are required for blocking the rapid apoptosis. In the third aim, we will compare the properties of the persistently infected cell with those of the acutely infected cells. For this purpose, the different steps of the viral life cycle will be examined. Moreover, the status of the innate immune response in the persistently infected cells will be evaluated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI073303-05
Application #
8242019
Study Section
Virology - B Study Section (VIRB)
Program Officer
Cassetti, Cristina
Project Start
2008-04-01
Project End
2013-08-31
Budget Start
2012-04-01
Budget End
2013-08-31
Support Year
5
Fiscal Year
2012
Total Cost
$346,221
Indirect Cost
$125,698
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Chattopadhyay, Saurabh; Sen, Ganes C (2014) Meet the terminator: The phosphatase PP2A puts brakes on IRF-3 activation. Mol Cell 54:210-1
Wetzel, Jaime L; Fensterl, Volker; Sen, Ganes C (2014) Sendai virus pathogenesis in mice is prevented by Ifit2 and exacerbated by interferon. J Virol 88:13593-601
Fensterl, Volker; Wetzel, Jaime L; Sen, Ganes C (2014) Interferon-induced protein Ifit2 protects mice from infection of the peripheral nervous system by vesicular stomatitis virus. J Virol 88:10303-11
Chattopadhyay, Saurabh; Sen, Ganes C (2014) dsRNA-activation of TLR3 and RLR signaling: gene induction-dependent and independent effects. J Interferon Cytokine Res 34:427-36
Chattopadhyay, Saurabh; Sen, Ganes C (2014) Tyrosine phosphorylation in Toll-like receptor signaling. Cytokine Growth Factor Rev 25:533-41
Chattopadhyay, Saurabh; Fensterl, Volker; Zhang, Ying et al. (2013) Role of interferon regulatory factor 3-mediated apoptosis in the establishment and maintenance of persistent infection by Sendai virus. J Virol 87:16-24
Yamashita, Michifumi; Millward, Carrie A; Inoshita, Hiroyuki et al. (2013) Antiviral innate immunity disturbs podocyte cell function. J Innate Immun 5:231-41
Chattopadhyay, Saurabh; Fensterl, Volker; Zhang, Ying et al. (2013) Inhibition of viral pathogenesis and promotion of the septic shock response to bacterial infection by IRF-3 are regulated by the acetylation and phosphorylation of its coactivators. MBio 4:
Leonova, Katerina I; Brodsky, Leonid; Lipchick, Brittany et al. (2013) p53 cooperates with DNA methylation and a suicidal interferon response to maintain epigenetic silencing of repeats and noncoding RNAs. Proc Natl Acad Sci U S A 110:E89-98
White, Christine L; Chattopadhyay, Saurabh; Sen, Ganes C (2011) Phosphatidylinositol 3-kinase signaling delays sendai virus-induced apoptosis by preventing XIAP degradation. J Virol 85:5224-7

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