The goal of the project is to investigate how a newly discovered antiviral pathway is activated and regulated and what its role is in determining viral pathogenesis and persistence. The type I interferon (IFN) system plays a major role in innate antiviral response. Virus infection activates the transcription factor IRF-3, which is responsible for the induction of IFN and other antiviral proteins. Recent investigation by us has revealed that IRF-3 also activates an IFN-independent pro-apoptotic pathway, named RIPA (RIG-I-activated IRF-3-mediated Pathway of Apoptosis). To trigger RIPA, activated IRF-3 binds Bax and translocates it to mitochondria to cause apoptosis;the two functions of IRF-3 are genetically separable. RIPA is activated by many RNA and DNA viruses and inhibits both viral replication and pathogenesis. RIPA is temporally regulated through the action of XIAP and PI3 kinase, which is also activated upon virus infection. In the absence of RIPA, viruses establish persistent infection. Here we propose to investigate RIPA further.
In Aim 1, we will investigate how IRF-3 is activated in RIPA as a consequence of phosphorylation of specific serine residues and ubiquitynation of specific lysine residues. Genetic and biochemical analyses will be used for this purpose.
In Aim 2, we will investigate how viruses evade RIPA temporarily, by triggering negative regulation of RIPA though activation of the epidermal growth factor receptor, which in turn activates PI3 kinase.
In Aim 3, we will investigate the physiological relevance of RIPA in determining the outcome of virus infection. Mutant cells defective in one, but not the other, action of IRF-3, will be used for measuring the role of RIPA on the efficiency of virus replication and establishing viral persistence. Finally, genetically modified mice, including new IRF-3 mutant knock-in mice which have been generated for this project, will be used to assess the in vivo contribution of RIPA in controlling viral pathogenesis.

Public Health Relevance

Because viral diseases are major health risks to the society, it is important to understand how they are caused and how our natural defense mechanisms can protect us. A virus-infected cell can be productively infected and produce a large number of infections progeny viruses;hence, an effective antiviral strategy, for the host, is to promote death of the infected cell before it produces more viruses. This proposal is to investigate how such suicidal response of the infected cell is triggered and regulated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI073303-06A1
Application #
8690329
Study Section
Virology - B Study Section (VIRB)
Program Officer
Cassetti, Cristina
Project Start
2007-04-01
Project End
2019-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
6
Fiscal Year
2014
Total Cost
$139,348
Indirect Cost
$51,431
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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