Jin, Jing
(2008)
REGULATION OF GAG TRAFFICKING DURING RETROVIRUS ASSEMBLY AND BUDDING.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
Retroviral Gag polyproteins are necessary and sufficient for virus budding, but little is known about how thousands of Gag polyproteins are transported to the budding sites. The actin cytoskeleton has long been speculated to take a role in retrovirus assembly and recent studies suggest that HIV-1 assembly is regulated as early as viral RNA nuclear export, however specific mechanisms for these regulations are unknown. In contrast to numerous studies of HIV-1 Gag assembly and budding, relatively little is reported for these fundamental pathways among animal lentiviruses. In this project, we used bimolecular fluorescence complementation (BiFC) (1) to reveal intimate (<15nm) and specific associations between EIAV Gag and actin, but not tubulin; (2) to characterize and compare assembly sites and budding efficiencies of EIAV and HIV-1 Gag in both human and rodent cells when the mRNA nuclear export context is altered to be Rev-dependent or Rev-independent; (3) to reveal co-assembly of Rev-dependent and Rev-independent HIV-1 Gag and rescued assembly of Rev-independent HIV-1 Gag in human cells by in cis provided membrane targeting signals. The results of these studies showed that (1) multimerization of EIAV Gag was required for association with filamentous actin and this association correlated with Gag budding efficiency, suggesting that association of Gag multimers with filamentous actin is important for efficient virion production; (2) HIV-1 and EIAV Gag assembled in different cellular at sites, and HIV-1 but not EIAV Gag assembly was affected by mRNA nuclear export pathways, suggesting that alternative cellular pathways can be adapted for lentiviral Gag assembly and budding; (3) Rev-independent HIV-1 Gag was deficient in lipid raft targeting and its assembly and budding could be restored by membrane targeting signals provided in trans or in cis, suggesting that raft association is critical for HIV-1 assembly and budding and is regulated as early as nuclear export of Gag-encoding mRNA. The findings presented in these studies are significant for public health because a better understanding of the mechanism of retrovirus assembly and budding increase the potential to develop novel antiviral therapies targeting this critical step in the viral life cycle.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
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| Date: |
30 January 2008 |
| Date Type: |
Completion |
| Defense Date: |
21 September 2007 |
| Approval Date: |
30 January 2008 |
| Submission Date: |
5 October 2007 |
| Access Restriction: |
5 year -- Restrict access to University of Pittsburgh for a period of 5 years. |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Public Health > Infectious Diseases and Microbiology |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
assembly and budding; retrovirus; BiFC; trafficking |
| Other ID: |
http://etd.library.pitt.edu/ETD/available/etd-10052007-153423/, etd-10052007-153423 |
| Date Deposited: |
10 Nov 2011 20:02 |
| Last Modified: |
15 Nov 2016 13:50 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/9437 |
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