Nucleic Acids Research, Vol 27, Issue 4 972-978, Copyright © 1999 by Oxford University Press
G Maga, M Amacker, U Hubscher, G Gosselin, JL Imbach, C Mathe, A Faraj, JP Sommadossi and S Spadari
In order to identify the basis for the relaxed enantio-selectivity of human
immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and to
evaluate possible cross-resistance patterns between L-nucleoside- ,
D-nucleoside- and non-nucleoside RT inhibitors, to be utilised in
anti-HIV-1 combination therapy, we applied an in vitro approach based on
the utilisation of six recom-binant HIV-1 RT mutants containing single
amino acid substitutions known to confer Nevirapine resistance in treated
patients. The mutants were compared on different RNA/DNA and DNA/DNA
substrates to the wild type (wt) enzyme for their sensitivity towards
inhibition by the D- and L-enantiomers of 2'-deoxy- and 2',3'-
dideoxynucleoside triphosphate analogs. The results showed that the 3'-
hydroxyl group of the L-(beta)-2'-deoxyribose moiety caused an unfavourable
steric hindrance with critic residues in the HIV-1 RT active site and this
steric barrier was increased by the Y181I mutation. Elimination of the
3'-hydroxyl group removed this hindrance and significantly improved binding
to the HIV-1 RT wt and to the mutants. These results demonstrate the
critical role of both the tyrosine 181 of RT and the 3'-position of the
sugar ring, in chiral discrimination between D- and L-nucleoside
triphosphates. Moreover, they provide an important rationale for the
combination of D- and L- (beta)-dideoxynucleoside analogs with
non-nucleoside RT inhibitors in anti-HIV chemotherapy, since
non-nucleosideinhibitors resistance mutations did not confer
cross-resistance to dideoxynucleoside analogs.
ARTICLES
Molecular basis for the enantioselectivity of HIV-1 reverse transcriptase: role of the 3'-hydroxyl group of the L-(beta)-ribose in chiral discrimination between D- and L-enantiomers of deoxy- and dideoxy-nucleoside triphosphate analogs
Institute of Biochemical and Evolutionary Genetics, National Research Council, I-27100, Pavia, Italy. maga@igbe.pv.cnr.it
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