Structure of HIV-1 reverse transcriptase with the inhibitor beta-Thujaplicinol bound at the RNase H active site.
Novel inhibitors are needed to counteract the rapid emergence of drug-resistant HIV variants. HIV-1 reverse transcriptase (RT) has both DNA polymerase and RNase H (RNH) enzymatic activities, but approved drugs that inhibit RT target the polymerase. Inhibitors that act against new targets, such as RNH, should be effective against all of the current drug-resistant variants. Here, we present 2.80 A and 2.04 A resolution crystal structures of an RNH inhibitor, beta-thujaplicinol, bound at the RNH active site of both HIV-1 RT and an isolated RNH domain. beta-thujaplicinol chelates two divalent metal ions at the RNH active site. We provide biochemical evidence that beta-thujaplicinol is a slow-binding RNH inhibitor with noncompetitive kinetics and suggest that it forms a tropylium ion that interacts favorably with RT and the RNA:DNA substrate.
Himmel
Daniel M
DM
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
Maegley
Karen A
KA
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Pauly
Tom A
TA
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Bauman
Joseph D
JD
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
Das
Kalyan
K
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
Dharia
Chhaya
C
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
Clark
Arthur D
AD
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
Ryan
Kevin
K
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Hickey
Michael J
MJ
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Love
Robert A
RA
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Hughes
Stephen H
SH
HIV Drug Resistance Program, NCI-Frederick, Frederick, MD 21702-1201, USA.
Bergqvist
Simon
S
Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
Arnold
Eddy
E
Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA. Electronic address: arnold@cabm.rutgers.edu.
D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D054303:HIV Reverse Transcriptase / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism*; D014334:Tropolone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
D018894:Reverse Transcriptase Inhibitors; C497842:beta-thujaplicinol; D014334:Tropolone; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D012260:Ribonucleases