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The structure of the human tRNALys3 anticodon bound to the HIV genome Loop I is stabilized by modified nucleosides
Author(s) -
Bilbille Yann,
Vendeix Franck AP,
Guenther Richard,
Malkiewicz Andrzej,
Ariza Xavier,
Vilarrasa Jaume,
Agris Paul F
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.845.1
Subject(s) - transfer rna , base pair , duplex (building) , primer binding site , rna , reverse transcriptase , pseudouridine , primer (cosmetics) , biology , chemistry , genetics , dna , gene , organic chemistry
Replication of HIV requires base pairing of the reverse transcriptase primer, human tRNA Lys3 , to the viral RNA. Although the major complementary base pairing occurs between the HIV primer binding sequence and the tRNA's 3′‐terminus, an important discriminatory, secondary contact occurs between the viral A‐rich Loop I and the modified, U‐rich anticodon domain of tRNA Lys3 . The thermal stabilities of variously modified tRNA anticodon region sequences bound to the Loop I of viral serosubtypes G and B were analyzed by UV and CD and the structure of one duplex was determined using NMR spectroscopy and restrained molecular dynamics. Thiouridine, s 2 U 34 , and pseudouridine, ? 39 , modifications appreciably stabilized the interaction of the anticodon region with the viral RNAs. The structure of the duplex composed of the HIV‐1 Loop I bound to a doubly modified tRNA's anticodon region resulted in two coaxially stacked A‐form RNA stems separated by two mismatched base pairs, U?? and G?A, that maintained a reasonable helix diameter. Thus, the important interaction of the primer's anticodon region with the viral RNA is strengthened by s 2 U 34 and ? 39 modifications that facilitate duplex interaction. This research was supported by a National Institutes of Health grant and National Science Foundation grants.