Preferences for the selection of unique tRNA primers revealed from analysis of HIV-1 replication in peripheral blood mononuclear cells

Kenda L Moore-Rigdon,Casey D Morrow,Barry R Kosloff,Richard L Kirkman

doi:10.1186/1742-4690-2-21

Kenda L Moore-Rigdon, Casey D Morrow + Show 2 more

Open Access

https://doi.org/10.1186/1742-4690-2-21

Copy DOI

Journal: Retrovirology	Publication Date: Mar 24, 2005
Citations: 32	License type: cc-by

Affiliation: University of Alabama at Birmingham

Abstract

BackgroundAll human immunodeficiency virus (HIV-1) uses a host tRNALys,3 as the primer for reverse transcription. The tRNALys,3 is bound to a region on the HIV-1 genome, the primer-binding site (PBS), that is complementary to the 18 terminal nucleotides of tRNALys,3. How HIV-1 selects the tRNA from the intracellular milieu is unresolved.ResultsHIV-1 tRNA primer selection has been investigated using viruses in which the primer-binding site (PBS) and a sequence within U5 were altered so as to be complementary to tRNAMet, tRNAPro or tRNAIle. Analysis of the replication of these viruses in human peripheral blood mononuclear cells (PBMC) revealed preferences for the selection of certain tRNAs. HIV-1 with the PBS altered to be complementary to tRNAMet, with and without the additional mutation in U5 to be complementary to the anticodon of tRNAMet, stably maintains the PBS complementary to tRNAMet following extended in vitro culture in PBMC. In contrast, viruses with either the PBS or PBS and U5 mutated to be complementary to tRNAIle were unstable during in vitro replication in PBMC and reverted to utilize tRNALys,3. Viruses with the PBS altered to be complementary to tRNAPro replicated in PBMC but reverted to use tRNALys,3; viruses with mutations in both the U5 and PBS complementary to tRNAPro maintained this PBS, yet replicated poorly in PBMC.ConclusionThe results of these studies demonstrate that HIV-1 has preferences for selection of certain tRNAs for high-level replication in PBMC.

Highlights

All human immunodeficiency virus (HIV-1) uses a host tRNALys,3 as the primer for reverse transcription
Construction and characterization of Human immunodeficiency virus type 1 (HIV-1) proviral genomes complementary to tRNAMet, tRNAPro and tRNAIle In previous studies, we have described the construction of HIV-1 proviral genomes in which the primer-binding site (PBS) was made complementary to alternative tRNAs [15,16,17,18,19]
This result highlights that different tRNAs are available in peripheral blood mononuclear cells (PBMC) for capture by HIV-1 for use as the primer for reverse transcription

Summary

Introduction

All human immunodeficiency virus (HIV-1) uses a host tRNALys, as the primer for reverse transcription. The major steps in reverse transcription have been known for some time, the process by which the tRNA primer is selected from the intracellular milieu by the virus is less well understood. Human immunodeficiency virus type 1 (HIV-1), as do all lentiviruses, selects tRNALys, for use as the primer for reverse (page number not for citation purposes). How and why HIV-1 exclusively selects tRNALys, as the primer for reverse transcription is not known. Additional studies have shown that the specific incorporation of lysyl tRNA synthetase and its interaction with Gag could be important for the specific capture of tRNALys, used for initiation of reverse transcription [9,10,11]

Methods

Results

Conclusion