Circularization of the HIV-1 genome facilitates strand transfer during reverse transcription.
Two obligatory DNA strand transfers take place during reverse transcription of a retroviral RNA genome. The first strand transfer involves a jump from the 5' to the 3' terminal repeat (R) region positioned at each end of the viral genome. The process depends on base pairing between the cDNA synthesized from the 5' R region and the 3' R RNA. The tertiary conformation of the viral RNA genome may facilitate strand transfer by juxtaposing the 5' R and 3' R sequences that are 9 kb apart in the linear sequence. In this study, RNA sequences involved in an interaction between the 5' and 3' ends of the HIV-1 genome were mapped by mutational analysis. This interaction appears to be mediated mainly by a sequence in the extreme 3' end of the viral genome and in the gag open reading frame. Mutation of 3' R sequences was found to inhibit the 5'-3' interaction, which could be restored by a complementary mutation in the 5' gag region. Furthermore, we find that circularization of the HIV-1 genome does not affect the initiation of reverse transcription, but stimulates the first strand transfer during reverse transcription in vitro, underscoring the functional importance of the interaction.
Beerens
Nancy
N
Department of Molecular Biology, Aarhus University, DK-8000 Aarhus C, Denmark.
Kjems
Jørgen
J
D020413:3' Untranslated Regions / Q000235:genetics; D020121:5' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D018014:Gene Transfer Techniques; D016679:Genome, Viral; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry / Q000235:genetics*; D000079962:RNA, Circular; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D018626:Retroelements / Q000235:genetics; D014158:Transcription, Genetic; D014779:Virus Replication / Q000235:genetics
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D000079962:RNA, Circular; D012367:RNA, Viral; D018626:Retroelements; D012313:RNA; D012194:RNA-Directed DNA Polymerase