The long term objective of this proposal is to determine how poliovirus, the prototype agent of a medically important group of viruses (picornaviruses), inhibits initiation of host cell RNA synthesis by cellular RNA polymerases I, II and III. Previous studies have identified four sequence specific DNA binding Pol II transcription factors (TBP, the TATA binding protein, CREB, the cyclic AMP-responsive element binding protein, Oct-1, the octamer binding factor, and transcriptional activator p53), one Pol III factor, TFIIIC, which interacts with Pol III promoter, and SL-1, a Pol I factor, all of which are transcriptionally inactivated in virus-infected cells. Both biochemical and genetic evidence suggests that the virus-encoded protease, 3Cpro, cleaves these factors in vivo and in vitro and is directly responsible for host cell transcription shut-off. Biochemical, serological and genetic approaches will be used to determine the mechanism(s) of inactivation of transcription factors in virus-infected cells. Specifically, how TBP and Oct-1 cleavage by 3Cpro contribute to general Pol II and small nuclear RNA (snRNA) transcription will be addressed. Because TBP is also required of Pol III and Pol I transcription (in addition to Pol II transcription), experiments are proposed to elucidate the role of TBP cleavage in Pol III-catalyzed snRNA transcription and Pol I transcription. Additionally, the mechanism by which cleavage of one of the four subunits (TAF110, TBP associated factor) of the Pol I factor SL-1, contribute to Pol I shut-off, will be addressed. The mechanism of nuclear entry of the viral protease (3Cpro) required to shut-off host cell transcription will be studied using biochemical and genetic approaches. Finally, the mechanism by which a viral protein (2C) specifically stimulates Pol I transcription in vitro will be studied. Elucidation of the mechanism by which poliovirus negatively affects cellular transcription factor activities would undoubtedly facilitate a better understanding of virus-host interaction as well as regulation of transcription in eukaryotic cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI027451-11
Application #
2756585
Study Section
Virology Study Section (VR)
Program Officer
Meegan, James M
Project Start
1989-02-01
Project End
2004-01-31
Budget Start
1999-02-01
Budget End
2000-01-31
Support Year
11
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Banerjee, Rajeev; Weidman, Mary K; Navarro, Sonia et al. (2005) Modifications of both selectivity factor and upstream binding factor contribute to poliovirus-mediated inhibition of RNA polymerase I transcription. J Gen Virol 86:2315-22
Jhaveri, Ravi; Kundu, Pallob; Shapiro, Alan M et al. (2005) Effect of heptitis C virus core protein on cellular gene expression: specific inhibition of cyclooxygenase 2. J Infect Dis 191:1498-506
Kundu, Pallob; Raychaudhuri, Santanu; Tsai, Weimin et al. (2005) Shutoff of RNA polymerase II transcription by poliovirus involves 3C protease-mediated cleavage of the TATA-binding protein at an alternative site: incomplete shutoff of transcription interferes with efficient viral replication. J Virol 79:9702-13
Sharma, Rakhi; Raychaudhuri, Santanu; Dasgupta, Asim (2004) Nuclear entry of poliovirus protease-polymerase precursor 3CD: implications for host cell transcription shut-off. Virology 320:195-205
Banerjee, Rajeev; Weidman, Mary K; Echeverri, Angela et al. (2004) Regulation of poliovirus 3C protease by the 2C polypeptide. J Virol 78:9243-56
Weidman, Mary K; Sharma, Rahki; Raychaudhuri, Santanu et al. (2003) The interaction of cytoplasmic RNA viruses with the nucleus. Virus Res 95:75-85
Banerjee, R; Dasgupta, A (2001) Interaction of picornavirus 2C polypeptide with the viral negative-strand RNA. J Gen Virol 82:2621-7
Banerjee, R; Tsai, W; Kim, W et al. (2001) Interaction of poliovirus-encoded 2C/2BC polypeptides with the 3' terminus negative-strand cloverleaf requires an intact stem-loop b. Virology 280:41-51
Weidman, M K; Yalamanchili, P; Ng, B et al. (2001) Poliovirus 3C protease-mediated degradation of transcriptional activator p53 requires a cellular activity. Virology 291:260-71
Banerjee, R; Dasgupta, A (2001) Specific interaction of hepatitis C virus protease/helicase NS3 with the 3'-terminal sequences of viral positive- and negative-strand RNA. J Virol 75:1708-21

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