Unusual structure of the human immunodeficiency virus type 1 trans-activation response element.
The trans-activation response element (TAR) of human immunodeficiency virus type 1 is a structured RNA consisting of the first 60 nucleotides of all human immunodeficiency virus type 1 RNAs. Computer analyses and limited structural analyses indicated that TAR consists of a stem-bulge-loop structure. Mutational analyses showed that sequences in the bulge are required for Tat binding, whereas sequences in both the bulge and the loop are required for trans activation. In this study, we probed the structures of TAR and various mutants of TAR with chemical probes and RNases and used these methods to footprint a Tat peptide on TAR. Our data show that the structure of wild-type TAR is different from previously published models. The bulge, a Tat-binding site, consists of four nucleotides. The loop is structured, rather than simply single stranded, in a fashion reminiscent of the structures of the tetraloop 5'-UUCG-3' and the GNRA loop (C. Cheong, G. Varani, and I. Tinoco, Jr., Nature [London] 346:680-682, 1990; H.A. Heus and A. Pardi, Science 253:191-193, 1991). RNA footprint data indicate that three bases in the bulge are protected and suggest that a conformational change occurs upon Tat binding.
Colvin
R A
RA
Section of Cell Growth, Regulation and Oncogenesis, Duke University Medical Center, Durham, North Carolina 27710.
Garcia-Blanco
M A
MA
D001483:Base Sequence; D001665:Binding Sites; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D008961:Models, Structural; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D000099253:Trans-Activation Responsive RNA-Binding Protein
D009838:Oligodeoxyribonucleotides; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D000099253:Trans-Activation Responsive RNA-Binding Protein