HIV-1 rev regulation involves recognition of non-Watson-Crick base pairs in viral RNA

Bartel, David P.; Zapp, Maria L.; Green, Michael R.; Szostak, Jack W.

doi:10.1016/0092-8674(91)90527-6

Cited by 380 publications

(373 citation statements)

References 39 publications

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“…Systematic evolution of ligands by exponential enrichment (SELEX) is an in vitro selection and evolution method that has been used to isolate nucleic acid polymers that bind to various molecules (12,13). The SELEX process and similar methods have been used previously to isolate RNA ligands that bind to the HIV-1 rev (14)(15)(16)(17), tat (18), reverse transcriptase (19,20) and integrase (21) proteins. Another screening method has been used to isolate nucleic acids that bind to the HIV-1 envelope protein (22).…”

Section: Introductionmentioning

confidence: 99%

In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein

Lochrie

Waugh²,

Pratt³

et al. 1997

Nucleic Acids Research

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Section: Introductionmentioning

confidence: 99%

In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein

Lochrie

Waugh²,

Pratt³

et al. 1997

Nucleic Acids Research

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“…The only other attempt to delineate a protein binding site by in vitro selection from a partially randomized pool of sequences was made with the Rev protein (24). In that case, a very high rate of mutagenesis (65% wild type at any one position) was applied to a 66 nucleotide domain and only two short segments, forming an internal loop, showed a much reduced level of mutagenesis.…”

Section: Discussionmentioning

confidence: 99%

On the role of rRNA tertiary structure in recognition of ribosomal protein L11 and thiostrepton

Draper

1995

Nucl Acids Res

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“…After 1 hr at 37°C, the binding reactions were filtered over nitrocellulose filters, and the bound RNA was eluted and loaded onto a denaturing polyacrylamide gel as modified from ref. 23. The binding ratio of individual aptamers was measured analogously.…”

Section: Methodsmentioning

confidence: 99%

“…The RNA was ethanol precipitated in the presence of 20 g glycogen, redissolved in TE buffer, and reverse transcribed; cDNA was PCR amplified, followed by in vitro transcription. Cycles 3 and 4 were performed in the presence of a 50-fold excess of wild-type RNA (23). After cycle 4, binding sequences were PCR amplified with primers 5 Ј-TCT AAT ACG ACT CAC TAT AGG GAG GAT CCC GCT CGT GTC-3 Ј and 5Ј-GCT TGA ATT CGT AAT GCT CAT TGC-3 Ј, restriction digested with BamHI and EcoRI, cloned, and sequenced.…”

Section: Methodsmentioning

confidence: 99%

In vitro and in vivo characterization of novel mRNA motifs that bind special elongation factor SelB

Klug

Hüttenhofer

Kromayer

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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The special elongation factor SelB of Escherichia coli promotes selenocysteine incorporation into formate dehydrogenases. This is thought to be achieved through simultaneous binding to selenocysteyl-tRNA Sec and, in the case of formate dehydrogenase H, to an fdhF mRNA hairpin structure 3 adjacent to the UGA selenocysteine codon. By in vitro selection, novel RNA sequences (''aptamers''), which can interact tightly and specifically with SelB, were isolated from an RNA library. The library was comprised of mutagenized variants of the wild-type fdhF mRNA hairpin. One-half of the selected sequences contained the apical stem-loop of the fdhF mRNA hairpin highly conserved. Some of the aptamers showed deviations in the primary sequence within this region of the wild-type fdhF hairpin motif while still binding with high affinity to SelB. Binding studies performed with truncated versions of SelB revealed that aptamers binding to different sites on the protein have been selected. To dissect SelB binding to the fdhF hairpin from the overall biological function of this complex, four selected aptamers were analyzed in vivo for UGA readthrough in a lacZ fusion construct. Among these, one promoted UGA readthrough in vivo. Three of the aptamers, however, were drastically reduced or unable to replace the fdhF mRNA hairpin in vivo, despite the similar secondary structure and binding affinities of these RNAs compared with the wild-type motif. This finding implies functions of the fdhF hairpin that go beyond the mere tethering of selenocysteyl-tRNA Sec to the UGA codon.

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HIV-1 rev regulation involves recognition of non-Watson-Crick base pairs in viral RNA

Cited by 380 publications

References 39 publications

In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein

In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein

On the role of rRNA tertiary structure in recognition of ribosomal protein L11 and thiostrepton

In vitro and in vivo characterization of novel mRNA motifs that bind special elongation factor SelB

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