Crystal structure of an eight-base pair duplex containing the 3'-DNA-RNA-5' junction formed during initiation of minus-strand synthesis of HIV replication.
During initiation of minus-strand synthesis by HIV-1 reverse transcriptase, a 3'-DNA-RNA-5' junction is formed involving the 3'-end of tRNAlys,3. The HIV-RT-associated RNase H cleaves the RNA template strand specifically, opposite the newly synthesized DNA strand. We have determined the crystal structure at 1.9 A resolution of an eight-base pair hybrid duplex representing the junction to identify global or local structural perturbations which may be recognized by HIV-RT RNase H. The junction octamer is in a global A-type conformation throughout. A base pair step with distinct stacking geometry and variable backbone conformation is located next to the main endonucleolytic cleavage site. This base pair step may serve as a recognition site for HIV-RT RNase H.
Forschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
U
U
Mueller
G
G
Maier
A
A
Mochi Onori
L
L
Cellai
H
H
Heumann
U
U
Heinemann
D001482:Base Composition; D003460:Crystallization; D018360:Crystallography, X-Ray; D004279:DNA, Viral / Q000096:biosynthesis / Q000737:chemistry*; D054303:HIV Reverse Transcriptase / Q000737:chemistry; D015497:HIV-1 / Q000235:genetics*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009692:Nucleic Acid Heteroduplexes / Q000096:biosynthesis / Q000737:chemistry*; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis / Q000737:chemistry*; D016914:Ribonuclease H / Q000737:chemistry; D014779:Virus Replication / Q000235:genetics*
D004279:DNA, Viral; D009692:Nucleic Acid Heteroduplexes; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D054303:HIV Reverse Transcriptase; D016914:Ribonuclease H