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J Mol Biol. 2011 Jul 29;410(5):863-74. doi: 10.1016/j.jmb.2011.04.024.

Secondary structure of the HIV reverse transcription initiation complex by NMR.

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1
Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA. epuglisi@stanford.edu

Abstract

Initiation of reverse transcription of genomic RNA is a key early step in replication of the human immunodeficiency virus (HIV) upon infection of a host cell. Viral reverse transcriptase initiates from a specific RNA-RNA complex formed between a host transfer RNA (tRNA(Lys)(3)) and a region at the 5' end of genomic RNA; the 3' end of the tRNA acts as a primer for reverse transcription of genomic RNA. We report here the secondary structure of the HIV genomic RNA-human tRNA(Lys)(3) initiation complex using heteronuclear nuclear magnetic resonance methods. We show that both RNAs undergo large-scale conformational changes upon complex formation. Formation of the 18-bp primer helix with the 3' end of tRNA(Lys)(3) drives large conformational rearrangements of the tRNA at the 5' end while maintaining the anticodon loop for potential loop-loop interactions. HIV RNA forms an intramolecular helix adjacent to the intermolecular primer helix. This helix, which must be broken by reverse transcription, likely acts as a kinetic block to reverse transcription.

PMID:
21763492
PMCID:
PMC3710119
DOI:
10.1016/j.jmb.2011.04.024
[Indexed for MEDLINE]
Free PMC Article
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