Nucleic Acids Research, Vol 24, Issue 19 3733-3738, Copyright © 1996 by Oxford University Press
PR Bohjanen, RA Colvin, M Puttaraju, MD Been and MA Garcia-Blanco
Linear TAR RNA has previously been used as a decoy to inhibit HIV-1
transcription in vitro and HIV-1 replication in vivo. A 48 nucleotide
circular RNA containing the stem, bulge and loop of the HIV-1 TAR element
was synthesized using the self-splicing activity of a group I permuted
intron-exon and was tested for its ability to function as a TAR decoy in
vitro. This small circular TAR molecule was exceptionally stable in HeLa
nuclear extracts, whereas a similar linear TAR molecule was rapidly
degraded. The TAR circle bound specifically to Tfr38, a peptide containing
the TAR-binding region of Tat. The ability of Tat to trans-activate
transcription from the HIV-1 promoter in vitro was efficiently inhibited by
circular TAR RNA but not by TAR circles that contained either bulge or loop
mutations. TAR circles did not inhibit transactivation exclusively by
binding to Tat since this inhibition was not reversed by adding excess Tat
to the transcription reaction. Together, these data suggest that TAR
circles act as decoys that inhibit transactivation by binding to Tat and at
least one cellular factor. These data also demonstrate the utility of small
circular RNA molecules as tools for biochemical studies.
ARTICLES
A small circular TAR RNA decoy specifically inhibits Tat-activated HIV- 1 transcription
Division of Infectious Diseases, Duke University Medical Center, Durham, NC 27710, USA.
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