Nucleic Acids Research, 1992, Vol. 20, No. 15 3945-3953
© 1992
MOLECULAR BIOLOGY |
Enhancement of ribosomal frameshifting by oligonucleotides targeted to the HIV gag-polregion
ISIS Pharmaceuticals 2280 Faraday Avenue, Carlsbadm CA 92008, USA
*To whom Correspondence should be addressed
Received May 14, 1992. Revised June 30, 1992. Accepted June 30, 1992.
The polgene of all retroviruses is expressed as a gagpol fusion protein which is proteolytically processed to produce all viral enzymes. In the human immunodeficiency virus (HIV), the gag and pol genes overlap by 241 nucleotides with pol in the - 1 phase with respect to gag. The gag-pol fusion is produced via a - 1 ribosomal frameshifting event that brings the overlapping, out-of-phase gag and pol genes into translational phase. Frameshifting occurs at a so called ‘shift site’ 8 – 10 nucleotides upstream of a hairpin loop which may play a role in the regulation of frameshifting. We have fused this region of HIV-1 to the 5' end of the firefly luciferase reporter gene in order to quantitatively measure ribosomal frameshifting both in cells and by in vitro translation. A series of 2'-O-methyl oligonucleotides was designed to specifically bind the sequences which flank the gag-pol hairpin. Ribosomal frameshifting is enhanced up to 6 fold by those oligonucleotides which bind the area just 3 to the stem. Oligonucleotides which bind 5' to the stem have no effect on framshift efficiency. In addition, we have constructed a series of fusion genes which mimic the effect of the bound oligonucleotides with intramolecular hairpins. The results suggest that increasing RNA secondary structure downstream of the shift site increases the frequency of ribosomal frameshifting, and that this effect can be mimicked by antlsense oligonucleotides.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. T. HOWARD, R. F. GESTELAND, and J. F. ATKINS Efficient stimulation of site-specific ribosome frameshifting by antisense oligonucleotides RNA, October 20, 2004; 10(10): 1653 - 1661. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Sei, Q.-e. Yang, D. O'Neill, K. Yoshimura, K. Nagashima, and H. Mitsuya Identification of a Key Target Sequence To Block Human Immunodeficiency Virus Type 1 Replication within the gag-pol Transframe Domain J. Virol., May 15, 2000; 74(10): 4621 - 4633. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. Aupeix-Scheidler, S. Chabas, L. Bidou, J.-P. Rousset, M. Leng, and J.-J. Toulme Inhibition of in vitro and ex vivo translation by a transplatin-modified oligo(2'-O-methylribonucleotide) directed against the HIV-1 gag-pol frameshift signal Nucleic Acids Res., January 15, 2000; 28(2): 438 - 445. [Abstract] [Full Text] [PDF]
|
|