Inhibition of HIV derived lentiviral production by TAR RNA binding domain of TAT protein.
BACKGROUND
A 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.


RESULTS
We 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.


CONCLUSIONS
Tat-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.
Mi
Michael Y
MY
Department of Dermatology and Immunology, University of Pittsburgh, School of Medicine, Pittsburgh, PA 15261, USA. mikemi235@gmail.com
Zhang
Jiying
J
He
Yukai
Y
D019380:Anti-HIV Agents / Q000494:pharmacology*; D001665:Binding Sites; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D015696:Gene Products, tat / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006678:HIV / Q000187:drug effects / Q000502:physiology; D016325:HIV Long Terminal Repeat; D006801:Humans; D008854:Microscopy, Electron; D010446:Peptide Fragments / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D012313:RNA / Q000378:metabolism*; D014161:Transduction, Genetic; D014779:Virus Replication / Q000187:drug effects*; D054322:tat Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D015696:Gene Products, tat; D010446:Peptide Fragments; D054322:tat Gene Products, Human Immunodeficiency Virus; D012313:RNA