The long-term goal of the research described in this proposal is to elucidate fundamental aspects of HIV virion formation and RNA packaging. Although the outline of the process of assembly has been known for some time, and closely follows that of the simpler retroviruses, many unanswered questions remain. Elucidating the mechanism of HIV viral particle formation and RNA packaging in more detail is likely to help identify key events that could become targets for the next generation of anti-HIV therapeutics. The applicant will address the following specific questions:
Aim #1 : Do assembly and RNA packaging interactions begin on the polyribosome? The applicant will explore this question by expressing Gagpol together with Gag, and independently from it on a different mRNA, to determine whether sythesis of Gagpol by frameshifting contributes to its incorporation into particles. The applicant will also address the issue of cis versus trans packaging of viral RNA.
Aim #2 : Do RNA localization and processing mechanisms contribute to HIV RNA packaging? Although specific packaging signals are known to exist within HIV genomic RNA, and the NC protien seems to play role in RNA recognition, little is known about other factors that facilitate packaging. In this aim the applicant will: a) determine if poly A plays a role in RNA packaging; b) determine if targeting an RNA to the surface of the endoplasmic reticulum affects its ability to be packaged; c) determine if the pathway of nuclear export affects packaging efficiency; d) determine if newly identified RNA trafficking signals (RTS) present in the HIV genome and """"""""zipcodes"""""""" influence RNA packaging or assembly.
Aim #3 : What is the role of the 5' TAR element in RNA packaging and dimer linkage formation? When is the dimer linkage formed? The applicant has previously shown that the bottom part of the TAR stem is a necessary structural element for efficient RNA packaging. In this aim he will further explore this requirement, focusing attention on the possible role of TAR in dimer formation and dimer stability. He will also address the temporal order of dimer formation, i.e. can an RNA package without forming a dimer?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI047008-03
Application #
6511218
Study Section
Special Emphasis Panel (ZRG1-AARR-1 (01))
Program Officer
Young, Janet M
Project Start
2000-04-15
Project End
2004-03-31
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
3
Fiscal Year
2002
Total Cost
$257,702
Indirect Cost
Name
University of Virginia
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Olivieri, Kevin C; Scoggins, Robert M; Broderick, Brooks et al. (2008) Nef does not contribute to replication differences between R5 pre-AIDS and AIDS HIV-1 clones from patient ACH142. Retrovirology 5:42
Olivieri, Kevin; Scoggins, Robert M; Bor, Yeou-cherng et al. (2007) The envelope gene is a cytopathic determinant of CCR5 tropic HIV-1. Virology 358:23-38
Swartz, Jennifer E; Bor, Yeou-Cherng; Misawa, Yukiko et al. (2007) The shuttling SR protein 9G8 plays a role in translation of unspliced mRNA containing a constitutive transport element. J Biol Chem 282:19844-53
Bor, Yeou-cherng; Swartz, Jennifer; Morrison, Avril et al. (2006) The Wilms' tumor 1 (WT1) gene (+KTS isoform) functions with a CTE to enhance translation from an unspliced RNA with a retained intron. Genes Dev 20:1597-608
Alexander, Melissa; Bor, Yeou-cherng; Ravichandran, Kodimangalam S et al. (2004) Human immunodeficiency virus type 1 Nef associates with lipid rafts to downmodulate cell surface CD4 and class I major histocompatibility complex expression and to increase viral infectivity. J Virol 78:1685-96
Coyle, John H; Guzik, Brian W; Bor, Yeou-Cherng et al. (2003) Sam68 enhances the cytoplasmic utilization of intron-containing RNA and is functionally regulated by the nuclear kinase Sik/BRK. Mol Cell Biol 23:92-103
Jin, Li; Guzik, Brian W; Bor, Yeou-cherng et al. (2003) Tap and NXT promote translation of unspliced mRNA. Genes Dev 17:3075-86
Levesque, L; Guzik, B; Guan, T et al. (2001) RNA export mediated by tap involves NXT1-dependent interactions with the nuclear pore complex. J Biol Chem 276:44953-62
Mouland, A J; Xu, H; Cui, H et al. (2001) RNA trafficking signals in human immunodeficiency virus type 1. Mol Cell Biol 21:2133-43
Guzik, B W; Levesque, L; Prasad, S et al. (2001) NXT1 (p15) is a crucial cellular cofactor in TAP-dependent export of intron-containing RNA in mammalian cells. Mol Cell Biol 21:2545-54