The investigator proposes to define sequences in the HIV env gene that are responsive to host cell control. This will be attempted by deleting sequences in the env gene and determining the effects of these deletions on viral RNA localization. In addition, the investigator proposes to add sequences from env and gag (specifies virion core proteins) genes to heterologous genes. After defining minimal sequences that prevent cytoplasmic accumulation of viral RNA in COS cells, the applicant will try to determine directly whether or not there is a role for these sequences on the efficiency of RNA splicing and whether or not there is a role for these sequences on RNA stability. Another aspect of this project is to try to characterize the mechanism by which env protein production is suppressed in the absence of Rev and to determine how Rev overcomes this suppression in HeLa cells. Additionally, the investigator proposes to determine the differences between the regulation of env RNA in COS and HeLa cells that account for differences in RNA localization in these two cell types. Dr. Emerman also will try to test a variety of other cell types for their regulation of env RNA localization. The applicant has developed a novel cell line for determination of viral titers. This cell line will be used to determine the effects of over-expression of each of the HIV regulatory genes on virus growth in lymphocytes. Quantification of viral spread through these cell lines may allow the determination of which steps of the virus life cycle are limiting and whether or not replication of virus can be hindered by inappropriate temporal expression of its regulatory genes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI030927-02
Application #
3145938
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1991-08-01
Project End
1996-05-31
Budget Start
1992-08-01
Budget End
1993-05-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109
Garcia, Erin I; Emerman, Michael (2018) Recurrent Loss of APOBEC3H Activity during Primate Evolution. J Virol :
OhAinle, Molly; Helms, Louisa; Vermeire, Jolien et al. (2018) A virus-packageable CRISPR screen identifies host factors mediating interferon inhibition of HIV. Elife 7:
Roesch, Ferdinand; OhAinle, Molly; Emerman, Michael (2018) A CRISPR screen for factors regulating SAMHD1 degradation identifies IFITMs as potent inhibitors of lentiviral particle delivery. Retrovirology 15:26
Adolph, Madison B; Ara, Anjuman; Feng, Yuqing et al. (2017) Cytidine deaminase efficiency of the lentiviral viral restriction factor APOBEC3C correlates with dimerization. Nucleic Acids Res 45:3378-3394
Fregoso, Oliver I; Emerman, Michael (2016) Activation of the DNA Damage Response Is a Conserved Function of HIV-1 and HIV-2 Vpr That Is Independent of SLX4 Recruitment. MBio 7:
Wittkopp, Cristina J; Adolph, Madison B; Wu, Lily I et al. (2016) A Single Nucleotide Polymorphism in Human APOBEC3C Enhances Restriction of Lentiviruses. PLoS Pathog 12:e1005865
McLaughlin Jr, Richard N; Gable, Jacob T; Wittkopp, Cristina J et al. (2016) Conservation and Innovation of APOBEC3A Restriction Functions during Primate Evolution. Mol Biol Evol 33:1889-901
Mitchell, Patrick S; Young, Janet M; Emerman, Michael et al. (2015) Evolutionary Analyses Suggest a Function of MxB Immunity Proteins Beyond Lentivirus Restriction. PLoS Pathog 11:e1005304
Matsen 4th, Frederick A; Small, Christopher T; Soliven, Khanh et al. (2014) A novel Bayesian method for detection of APOBEC3-mediated hypermutation and its application to zoonotic transmission of simian foamy viruses. PLoS Comput Biol 10:e1003493
Refsland, Eric W; Hultquist, Judd F; Luengas, Elizabeth M et al. (2014) Natural polymorphisms in human APOBEC3H and HIV-1 Vif combine in primary T lymphocytes to affect viral G-to-A mutation levels and infectivity. PLoS Genet 10:e1004761

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