AEGiS-03CROI: Specificity and mechanism of HIV-1 inhibition by transdominant rev.

3rd Conference on Retroviruses and Opportunistic Infections

Washington, DC - January 28-February 1, 1996

SPECIFICITY AND MECHANISM OF HIV-1 INHIBITION BY TRANSDOMINANT REV.

Conf Retroviruses Opportunistic Infect 1996 Jan 28-Feb 1; 3rd:57 (abstract no. 23)

Pavlakis GN1, Stauber R¹, Neumann M¹, Gaitanaris GA¹, Valentin A¹, Song S¹, Harrison J¹, Tabernero C², Felber BK²
¹ Human Retrovirus Section, ² Human Retrovirus Pathogenesis Group, ABL-BRP, NCI-FCRDC Frederick MD 21702-1201, USA

We have studied the function, localization and trafficking of the HIV-1 Rev protein and several Rev mutants by creating fusions with the autofluorescing Green fluorescent protein (GFP). This allowed the continuous observation of live human cells expressing Rev or the different Rev mutants alone or in combination. We showed that Rev-GFP protein fusions were functional. A transdominant Rev mutant (TDRev) fused to GFP (TDRev-GFP) was inhibiting specifically Rev function. Co-expression of Rev-GFP and TDRev showed that TDRev is responsible for retaining Rev in the nucleus, indicating that TDRev binds to and prevents the function of Rev. These results provide a mechanism for the function of TDRev.

We have previously identified cell lines that do not support efficient Rev function. Analysis of Rev trafficking in these cells indicated that the rates of Rev transport are severely affected, providing an explanation for the inefficient function of Rev.

In another line of experiments, we have shown that inhibition of HIV-1 expression by TDRev is specific and results solely from the inhibition of Rev function by TDRev. We have generated Rev-independent HIV-1 molecular clones able to propagate in human cells in the complete absence of Rev and its binding site, the RRE. These molecular clones are not inhibited in cell lines expressing TDRev, while the wild-type virus is. These results demonstrate the specificity of TDRev function against Rev.

^{Research sponsored by the National Cancer Institute, DHHS, undercontract with ABL.}

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Copyright © 1996 - Foundation for Retrovirology and Human Health . Reproduction of this abstract (other than one copy for personal reference) must be cleared through the Foundation for Retrovirology and Human Health.