AEGiS-14IAC: Relationship between tRNALys3 concentration, placement and viral infectivity in HIV-1.

14th International AIDS Conference

Barcelona, Spain - July 7-12, 2002

Relationship between tRNALys3 concentration, placement and viral infectivity in HIV-1.

Int Conf AIDS 2002 Jul 7-12; 14:(abstract no. A10016)

Niu MJ, Cen S, Gabor J, Javanbakht H, Wainberg MA, Kleiman L
Lady Davis Institute, Montreal, Canada

BACKGROUND: During the assembly of HIV-1, tRNALys3 is packaged into the virus, and is annealed to the virus genome where it serves as the primer for reverse transcription of minus strand DNA. In this work, we investigate whether there is any relationship between viral tRNALys3 content and annealing of tRNALys3 to the RNA genome or infectivity of virus.

METHODS: Viruses were isolated from COS7 cells were transfected with SVC21BH10, SVC21BH10-Lys3 and SVC21BH10Lys1,2, which contain both wild-type HIV-1 proviral DNA and a human tRNALys3 or tRNALys1,2 gene, respectively. Total viral RNA and virion were analyzed by dot blot, 2D PAGE, primer extension and infectivity assay.

RESULTS: BH10-Lys3 has approximately 1.6 times of packaged tRNALys3, 2 fold of genomic placement and infectivity of wild type, while BH10-Lys2 has less than one fifth the amount of tRNALys3 found in wild type virions, as well as less genomic placement of primer and infectivity.

CONCLUSION: The above results suggest that there is a direct correlation between the amount of tRNALys3 per copy of HIV-1 genomic RNA, the priming ability of the viral RNA sample and the infectivity of the virus.

Keywords: AEGIS, HIV-1, RNA, Viral, HIV-1 Reverse Transcriptase, Virion, RNA, Transcription, Genetic, Oligonucleotides, Antisense, Human, genetics, virology

020707
A10016