This proposal describes the genetic analysis of the Gag and Pol proteins of the Moloney murine leukemia virus and of the cellular host proteins with which they interact. We have used the yeast two-hybrid system to identify novel mammalian proteins that bind to various specific domains of the Gag and Pol gene products. We will now focus our ongoing efforts on several of these new targets: the IQGAP proteins, which interact with the MA portion of Gag; two components of the SUMO transferase system, which interact with the CA portion of Gag; and the ribosomal protein S3a, which binds the RNA pseudoknot at the Gag-Pol border. Biochemical methods will be used to confirm and characterize the interactions between the viral and cellular proteins both in vitro and in vivo. To determine the role of these proteins in virus replication, mutations in the viral genes will be identified that specifically abrogate the interaction with the host protein. These mutations will be introduced into the viral genome and the effects on virus replication in cell culture will be determined. Reversion analysis will be used to further define the requirements for the interaction, and to confirm the importance of the interaction for virus replication. Dominant interfering alleles and RNAi methods will be used to document the requirement for the interaction, and to define the stage of the life cycle at which the proteins act. The generality of the results will be tested by examining the effects of the genes on the replication of a panel of retroviruses of other families. These experiments will significantly extend our understanding of retrovirus replication, and may provide important new targets for antiviral intervention.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
3R37CA030488-25S2
Application #
7023512
Study Section
Virology Study Section (VR)
Program Officer
Ogunbiyi, Peter
Project Start
1981-08-01
Project End
2005-09-02
Budget Start
2005-02-01
Budget End
2005-09-02
Support Year
25
Fiscal Year
2005
Total Cost
$47,176
Indirect Cost
Name
Columbia University (N.Y.)
Department
Biochemistry
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Yang, Bin Xia; El Farran, Chadi A; Guo, Hong Chao et al. (2015) Systematic identification of factors for provirus silencing in embryonic stem cells. Cell 163:230-45
Schlesinger, Sharon; Meshorer, Eran; Goff, Stephen P (2014) Asynchronous transcriptional silencing of individual retroviral genomes in embryonic cells. Retrovirology 11:31
Wang, Gary Z; Wolf, Daniel; Goff, Stephen P (2014) EBP1, a novel host factor involved in primer binding site-dependent restriction of moloney murine leukemia virus in embryonic cells. J Virol 88:1825-9
Schlesinger, Sharon; Goff, Stephen P (2013) Silencing of proviruses in embryonic cells: efficiency, stability and chromatin modifications. EMBO Rep 14:73-9
Schlesinger, Sharon; Lee, Andreia H; Wang, Gary Z et al. (2013) Proviral silencing in embryonic cells is regulated by Yin Yang 1. Cell Rep 4:50-8
Zhu, Yiping; Wang, Xinlu; Goff, Stephen P et al. (2012) Translational repression precedes and is required for ZAP-mediated mRNA decay. EMBO J 31:4236-46
Arriagada, Gloria; Muntean, Lucia N; Goff, Stephen P (2011) SUMO-interacting motifs of human TRIM5? are important for antiviral activity. PLoS Pathog 7:e1002019
Houck-Loomis, Brian; Durney, Michael A; Salguero, Carolina et al. (2011) An equilibrium-dependent retroviral mRNA switch regulates translational recoding. Nature 480:561-4
Hogg, J Robert; Goff, Stephen P (2010) Upf1 senses 3'UTR length to potentiate mRNA decay. Cell 143:379-89
Rodriguez, Jason J; Goff, Stephen P (2010) Xenotropic murine leukemia virus-related virus establishes an efficient spreading infection and exhibits enhanced transcriptional activity in prostate carcinoma cells. J Virol 84:2556-62

Showing the most recent 10 out of 82 publications