Inhibition of HIV-1 replication using a mutated tRNALys-3 primer.
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.
Lu
Y
Y
UCLA Department of Medicine, Division of Hematology-Oncology, UCLA AIDS Institute, Los Angeles, California 90095, USA.
Planelles
V
V
Li
X
X
Palaniappan
C
C
Day
B
B
Challita-Eid
P
P
Amado
R
R
Stephens
D
D
Kohn
D B
DB
Bakker
A
A
Fay
P
P
Bambara
R A
RA
Rosenblatt
J D
JD
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
D017931:DNA Primers; D016655:HIV Core Protein p24; C034477:RNA primers; D012313:RNA; D054303:HIV Reverse Transcriptase