title: Inhibition of HIV-1 replication using a mutated tRNALys-3 primer.
abstract: Cellular tRNALys-3 serves as the primer for reverse transcription of human immunodeficiency virus, type 1 (HIV-1). tRNALys-3 interacts directly with HIV-1 reverse transcriptase, is packaged into viral particles and anneals to the primer-binding site (PBS) of the HIV-1 genome to initiate reverse transcription. Therefore, the priming step of reverse transcription is a potential target for antiviral strategies. We have developed a mutant tRNALys-3 derivative with mutations in the PBS-binding region such that priming specificity was re-directed to the highly conserved TAR stem-loop region. This mutant tRNA retains high-affinity binding to HIV-1 reverse transcriptase, viral encapsidation, and is able to prime at both the targeted TAR sequence and at the viral PBS. Constitutive expression of mutant tRNA in T-cells results in marked inhibition of HIV-1 replication, as determined by measurements of viral infectivity, syncytium formation, and p24 production. Inhibition of retroviral replication through interference with the normal process of priming constitutes a new anti-retroviral approach and also provides a novel tool for dissecting molecular aspects of priming.
authors.affiliation: UCLA Department of Medicine, Division of Hematology-Oncology, UCLA AIDS Institute, Los Angeles, California 90095, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Lu
authors.forename: V
authors.initials: V
authors.lastname: Planelles
authors.forename: X
authors.initials: X
authors.lastname: Li
authors.forename: C
authors.initials: C
authors.lastname: Palaniappan
authors.forename: B
authors.initials: B
authors.lastname: Day
authors.forename: P
authors.initials: P
authors.lastname: Challita-Eid
authors.forename: R
authors.initials: R
authors.lastname: Amado
authors.forename: D
authors.initials: D
authors.lastname: Stephens
authors.forename: D B
authors.initials: DB
authors.lastname: Kohn
authors.forename: A
authors.initials: A
authors.lastname: Bakker
authors.forename: P
authors.initials: P
authors.lastname: Fay
authors.forename: R A
authors.initials: RA
authors.lastname: Bambara
authors.forename: J D
authors.initials: JD
authors.lastname: Rosenblatt
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D015726:Giant Cells; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D012313:RNA; D013601:T-Lymphocytes / Q000821:virology; D014158:Transcription, Genetic; D014162:Transfection; D014771:Virion / Q000502:physiology; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D016655:HIV Core Protein p24; C034477:RNA primers; D012313:RNA; D054303:HIV Reverse Transcriptase