Biswas, Nabanita
(2011)
Adenosine Deaminase Acting on RNA (ADAR1) is a Novel Multitargeted Anti HIV-1 Cellular Protein.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
ADAR1 is an RNA editing enzyme which acts on completely or partially double-stranded RNA. Since HIV-1 RNA has such secondary structures, we have examined whether ADAR1 exhibits antiviral activity against HIV-1. Our results indicated that ADAR1 inhibited viral replication and infectious HIV-1 production in various cell lines including 293T, HeLa and Jurkat T cells, and was active against a number of X4- and R5-tropic HIV-1 of different clades. Analysis of the level of intracellular HIV-1 RNA showed no change in levels of intracellular gag, pol, and env RNA in the presence of ADAR1 despite a significant inhibition of intracellular and virion associated HIV-1 protein production. Furthermore mutational analysis showed that ADAR1 introduced most of the A-to-G mutations in the first exon of rev at positions 5998, 6011, 6017, and 6036 and in the Rev Response Element (RRE) binding region (positions 8413 and 8438) of rev and env RNA. In elucidating the mechanism of ADAR1 inhibition of HIV-1, we observed that A-G mutations in rev have a significant negative effect on the expression of Rev. However, all mutations could be complemented by wild type Rev.Furthermore, these A-G mutations in the RRE binding region of rev inhibited the binding of Rev to the RRE region in env and inhibited transport of primary transcripts like gag, pol and env from the nucleus to the cytoplasm. Introduction of these specific mutations in rev of an infectious molecular clone of HIV-1 by site directed mutagenesis abolished the replication capacity of HIV-1 by inhibiting viral protein synthesis without any effect on viral RNA synthesis, a phenotype exhibited by HIV-1-infected cells exposed to ADAR1. ADAR1 induced mutations in env further attenuated viral infectivity. ADAR1, thus, constitutes a novel class of cellular antiviral proteins with multiple targets in the viral genome thereby providing a new avenue of exploration for therapeutic drugs benefitting public health.
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Details
| Item Type: |
University of Pittsburgh ETD
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| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
23 September 2011 |
| Date Type: |
Completion |
| Defense Date: |
19 July 2011 |
| Approval Date: |
23 September 2011 |
| Submission Date: |
26 July 2011 |
| 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: |
ADAR1; Antiviral; Cellular Protein; HIV-1 |
| Other ID: |
http://etd.library.pitt.edu/ETD/available/etd-07262011-204453/, etd-07262011-204453 |
| Date Deposited: |
10 Nov 2011 19:54 |
| Last Modified: |
15 Nov 2016 13:47 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/8643 |
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