Aptamer displacement identifies alternative small-molecule target sites that escape viral resistance.
Aptamers targeting reverse transcriptase (RT) from HIV-1 inhibit viral replication in vitro, presumably by competing with binding of the primer/template complex. This site is not targeted by the currently available small-molecule anti-HIV-1 RT inhibitors. We have identified SY-3E4, a small-molecule inhibitor of HIV-1 RT, by applying a screening assay that utilizes a reporter-ribozyme regulated by the anti-HIV-1 RT aptamer. SY-3E4 displaces the aptamer from the protein, selectively inhibits DNA-dependent, but not RNA-dependent, polymerase activity, and inhibits the replication of both the wild-type virus and a multidrug-resistant strain. Analysis of available structural data of HIV-1 and HIV-2 RTs rationalizes many of the observed characteristics of the inhibitory profiles of SY-3E4 and the aptamer and suggests a previously not considered region in these RTs as a target for antiviral therapy. Our study reveals unexplored ways for rapidly identifying alternative small-molecule target sites in proteins and illustrates strategies for overcoming resistance-conferring mutations with small molecules.
Yamazaki
Satoko
S
LIMES Program Unit Chemical Biology & Medicinal Chemistry, c/o Kekulé Institute for Organic Chemistry & Biochemistry, University of Bonn, Gerhard-Domagk-Strasse 1, D-53121 Bonn, Germany.
Tan
Lu
L
Mayer
Günter
G
Hartig
Jörg S
JS
Song
Jin-Na
JN
Reuter
Sandra
S
Restle
Tobias
T
Laufer
Sandra D
SD
Grohmann
Dina
D
Kräusslich
Hans-Georg
HG
Bajorath
Jürgen
J
Famulok
Michael
M
D052157:Aptamers, Nucleotide; D002460:Cell Line; D024882:Drug Resistance, Viral; D054303:HIV Reverse Transcriptase / Q000737:chemistry / Q000235:genetics; D015497:HIV-1 / Q000201:enzymology / Q000502:physiology; D006801:Humans; D008958:Models, Molecular; D014779:Virus Replication
D052157:Aptamers, Nucleotide; D054303:HIV Reverse Transcriptase