title: Structure of HIV-1 reverse transcriptase with the inhibitor beta-Thujaplicinol bound at the RNase H active site.
abstract: 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.
authors.affiliation: Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Himmel
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Karen A
authors.initials: KA
authors.lastname: Maegley
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Tom A
authors.initials: TA
authors.lastname: Pauly
authors.affiliation: Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Bauman
authors.affiliation: Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
authors.forename: Kalyan
authors.initials: K
authors.lastname: Das
authors.affiliation: Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
authors.forename: Chhaya
authors.initials: C
authors.lastname: Dharia
authors.affiliation: Center for Advanced Biotechnology and Medicine and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854-8021, USA.
authors.forename: Arthur D
authors.initials: AD
authors.lastname: Clark
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Ryan
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Hickey
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Love
authors.affiliation: HIV Drug Resistance Program, NCI-Frederick, Frederick, MD 21702-1201, USA.
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Hughes
authors.affiliation: Pfizer Global Research and Development, La Jolla Laboratories, San Diego, CA 92121, USA.
authors.forename: Simon
authors.initials: S
authors.lastname: Bergqvist
authors.affiliation: 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.
authors.forename: Eddy
authors.initials: E
authors.lastname: Arnold
mesh_terms: 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
chemical_list: D018894:Reverse Transcriptase Inhibitors; C497842:beta-thujaplicinol; D014334:Tropolone; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D012260:Ribonucleases