Structural probing of the HIV-1 polypurine tract RNA:DNA hybrid using classic nucleic acid ligands.

Abstract:

:The interactions of archetypical nucleic acid ligands with the HIV-1 polypurine tract (PPT) RNA:DNA hybrid, as well as analogous DNA:DNA, RNA:RNA and swapped hybrid substrates, were used to probe structural features of the PPT that contribute to its specific recognition and processing by reverse transcriptase (RT). Results from intercalative and groove-binding ligands indicate that the wild-type PPT hybrid does not contain any strikingly unique groove geometries and/or stacking arrangements that might contribute to the specificity of its interaction with RT. In contrast, neomycin bound preferentially and selectively to the PPT near the 5'(rA)(4):(dT)(4) tract and the 3' PPT-U3 junction. Nuclear magnetic resonance data from a complex between HIV-1 RT and the PPT indicate RT contacts within the same regions highlighted on the PPT by neomycin. These observations, together with the fact that the sites are correctly spaced to allow interaction with residues in the ribonuclease H (RNase H) active site and thumb subdomain of the p66 RT subunit, suggest that despite the long cleft employed by RT to make contact with nucleic acids substrates, these sites provide discrete binding units working in concert to determine not only specific PPT recognition, but also its orientation on the hybrid structure.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Turner KB,Brinson RG,Yi-Brunozzi HY,Rausch JW,Miller JT,Le Grice SF,Marino JP,Fabris D

doi

10.1093/nar/gkn129

subject

Has Abstract

pub_date

2008-05-01 00:00:00

pages

2799-810

issue

8

eissn

0305-1048

issn

1362-4962

pii

gkn129

journal_volume

36

pub_type

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