Methods for analysis of incorporation and annealing methods for analysis of tRNA(Lys) in HIV-1

Shan Cen; Fei Guo; Lawrence Kleiman

doi:10.1007/978-1-59745-170-3_16

Methods for analysis of incorporation and annealing methods for analysis of tRNA(Lys) in HIV-1

Methods Mol Biol. 2009:485:223-32. doi: 10.1007/978-1-59745-170-3_16.

Authors

Shan Cen¹, Fei Guo, Lawrence Kleiman

Affiliation

¹ Department of Medicine, McGill University, Lady Davis Institute for Medical Research Jewish General Hospital, Montreal, QC, Canada.

PMID: 19020829
DOI: 10.1007/978-1-59745-170-3_16

Abstract

In HIV-1, tRNA(Lys3) serves as the primer for reverse transcription of minus strand strong stop cDNA. During viral assembly, the tRNA(Lys) isoacceptors, tRNA(Lys1,2) and tRNA(Lys3), are selectively packaged into the virion. The selectively packaging of tRNA(Lys3) facilitates the annealing of tRNA(Lys3) to the viral genome and the initiation of reverse transcription. We describe herein a set of experimental approaches for studying the mechanism by which tRNA(Lys) is selectively incorporated into HIV-1 and investigate how primer tRNA(Lys3) is annealed to viral genome. The methods described will also help in the analysis of cellular RNAs packaged in the virus particles.

MeSH terms

HIV-1 / physiology*
RNA, Transfer, Amino Acyl / genetics
RNA, Transfer, Amino Acyl / metabolism*
RNA, Viral / genetics
RNA, Viral / metabolism*
Virus Assembly*

Substances

RNA, Transfer, Amino Acyl
RNA, Viral