The long-term objective of the experiments described in this proposal is to increase our understanding of the replication of retroviruses, formerly called RNA tumor viruses. These agents, widely distributed in nature, cause a variety of leukemias, tumors and other diseases in animals and man. The specific goal of this work is to determine in as much detail as possible the structure and functions of the viral gene products encoded by the Moloney murine leukemia virus, a prototypical replication-competent retrovirus. Three major approaches will be taken toward that goal. First, mutagenesis of cloned DNA copies of the viral genome will be used to make specific alterations in selected regions, and the effects of these mutations on the viral life cycle will be determined by direct analyses of viral proteins and nucleic acids. Several domains of the viral gag, pol, and env genes, as well as various cis-acting regions, will be analyzed in this way. These experiments will determine the essential sequences in each gene or region, and will further define their roles in replication. Second, a major effort will be initiated to carry out reversion analysis of virus mutants. We will generate a series of replication-competent revertant viruses from several mutant parents, each arising by virtue of suppressing mutations that compensate for the effects of the original mutation. Mapping and analysis of these reversion mutations will help to locate domains of the viral gene products that interact with one another, and may help identify proteins that bind to viral DNA and RNA. Third, biochemical tests will be applied to three viral gene products expressed as fusion proteins in bacteria. Rapid assays will be developed to screen these proteins for various activities, such as DNA binding, tRNA binding, and nuclease activity. In each case, these tests will be applied to existing panels of mutants to help localize the regions needed for these activities.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37CA030488-13
Application #
3482190
Study Section
Virology Study Section (VR)
Project Start
1981-08-01
Project End
1995-01-31
Budget Start
1993-02-01
Budget End
1994-01-31
Support Year
13
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
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

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