Inhibition of HIV derived lentiviral production by TAR RNA binding domain of TAT proteinReport as inadecuate




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Retrovirology

, 2:71

First Online: 17 November 2005Received: 31 July 2005Accepted: 17 November 2005

Abstract

BackgroundA critical step in the production of new HIV virions involves the TAT protein binding to the TAR element. The TAT protein contains in close proximity its TAR RNA binding domain and protein transduction domain PTD. The PTD domain of TAT has been identified as being instrumental in the protein-s ability to cross mammalian cell and nuclear membranes. All together, this information led us to form the hypothesis that a protein containing the TAR RNA binding domain could compete with the native full length TAT protein and effectively block the TAR RNA binding site in transduced HIV infected cells.

ResultsWe synthesized a short peptide named Tat-P, which contained the TAR RNA binding and PTD domains to examine whether the peptide has the potential of inhibiting TAT dependent HIV replication. We investigated the inhibiting effects of Tat-P in vitro using a HIV derived lentiviral vector model. We found that the TAT PTD domain not only efficiently transduced test cells, but also effectively inhibited the production of lentiviral particles in a TAT dependent manner. These results were also supported by data derived from the TAT activated LTR-luciferase expression model and RNA binding assays.

ConclusionTat-P may become part of a category of anti-HIV drugs that competes with full length TAT proteins to inhibit HIV replication. In addition, this study indicates that the HIV derived lentiviral vector system is a safe and reliable screening method for anti-HIV drugs, especially for those targeting the interaction of TAT and TAR RNAs.

List of abbreviationsHIVHuman immunodeficiency virus

TARTrans-activating response region

TATTransactivating regulatory protein

PTDProtein transduction domain

RNARibonucleic acid

Tat-PTAT peptide

293TA human kidney epithelial cell line

Con-P1Control peptide one

Con-P2Control peptide two

EMElectron microscopy

PBSPhosphate buffered saline

TBSTris buffered saline

GFPGreen fluorescent protein

MTT3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide

HAARTHighly active antiretroviral therapy

ARVAnti-retroviral

FITCFluorescein isothiocyanate

VSV-GVesicular stomatitis virus glycoprotein

CMVCytomegalovirus

EMSAElectrophoretic mobility shift assay EMSA

Electronic supplementary materialThe online version of this article doi:10.1186-1742-4690-2-71 contains supplementary material, which is available to authorized users.

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Author: Michael Y Mi - Jiying Zhang - Yukai He

Source: https://link.springer.com/



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