Paradoxical production of target protein using antisense RNA expression vectors

Williard, Renee L.; Cc, Benz; Jd, Baxter; Kushner, Peter J.; Hunt, C. Anthony

doi:10.1016/0378-1119(94)90407-3

Cited by 8 publications

(2 citation statements)

References 19 publications

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“…Our results are in agreement with those of a previous study by Williard et al (36), in which ER activity was demonstrated to be significantly increased in cells expressing antisense ER RNA.…”

Section: Discussionsupporting

confidence: 94%

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Santagati

Gianazza

Agrati

et al. 1997

Molecular Endocrinology

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Antisense oligos complementary to the 5-end, but not to the 3-end, of the estrogen receptor (ER) messenger RNA caused a paradox accumulation of ER protein in MCF-7 cells. The same effect was observed after treatment of the cells with the corresponding sense oligos. The oligos interfering with ER down-regulation were demonstrated to specifically bind the ER with affinities in the nanomolar range. It is, therefore, proposed that the ER up-regulation induced by the oligos might be due to squelching of the ER (or ER-inducible proteins) from their binding site located in the 5-end of the ER gene. We also report that transcriptionally inactive ER mutants can undergo down-regulation, and that in denaturing gels, the migration profile of ER-oligo and ER-estrogen-responsive element complexes are dissimilar. We, therefore, propose that ER can interact with DNA in different ways and at different binding sites. These observations might have important pharmacological consequences, since specific drugs could be devised to induce the ER conformation necessary to perform only selected tasks of the ER transcriptional repertoire. (Molecular Endocrinology 11: 938-949, 1997)

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Section: Discussionsupporting

confidence: 94%

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Santagati

Gianazza

Agrati

et al. 1997

Molecular Endocrinology

View full text Add to dashboard Cite

show abstract

“…The present study stems from the observation of a paradoxical increase in ER content in MCF-7 cells treated with oligonucleotides aimed at blocking ER mRNA translation. Our results are in agreement with those of a previous study by Williard et al (36), in which ER activity was demonstrated to be significantly increased in cells expressing antisense ER RNA.…”

Section: Discussionsupporting

confidence: 94%

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Santagati

1997

Molecular Endocrinology

View full text Add to dashboard Cite

Antisense oligos complementary to the 5'-end, but not to the 3'-end, of the estrogen receptor (ER) messenger RNA caused a paradox accumulation of ER protein in MCF-7 cells. The same effect was observed after treatment of the cells with the corresponding sense oligos. The oligos interfering with ER down-regulation were demonstrated to specifically bind the ER with affinities in the nanomolar range. It is, therefore, proposed that the ER up-regulation induced by the oligos might be due to squelching of the ER (or ER-inducible proteins) from their binding site located in the 5'-end of the ER gene. We also report that transcriptionally inactive ER mutants can undergo down-regulation, and that in denaturing gels, the migration profile of ER-oligo and ER-estrogen-responsive element complexes are dissimilar. We, therefore, propose that ER can interact with DNA in different ways and at different binding sites. These observations might have important pharmacological consequences, since specific drugs could be devised to induce the ER conformation necessary to perform only selected tasks of the ER transcriptional repertoire.

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Comparison of three 3′ non-coding regions in Schizosaccharomyces pombe expression vectors: efficiencies of transcription termination and mRNA 3′-end formation

1996

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Analysis of an established Schizosaccharomyces pombe episomal shuttle vector suggested that inefficient transcription termination was deleterious to plasmid function. We undertook a study to determine if transcription in the presence and absence of 3'-processing within a vector could affect the ability of the plasmid to transform, transcribe and translate the RNA produced. This report provides an analysis of the effects that three S. pombe 3' non-coding regions have on the transformation and expression efficiencies of a fission yeast plasmid vector. The 3' regions from adh1, act1 and ura4 were tested for their ability to terminate and process adh1 promoter-driven transcription of a lacZ reporter gene. Differences between the 3'-processing sequences were observed, with transcription termination mediated by the ura4 3' region being more efficient than termination by the 3' regions of adh1 and act1. We show that plasmids containing inefficient transcription termination signals result in readthrough transcription and reduced transformation efficiencies. In addition, the readthrough transcripts containing 3' non-coding regions show impaired translation efficiencies. We describe an S. pombe vector (pURAS) with a high transformation efficiency that directs the production of highly translatable, discrete-sized transcripts.

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Paradoxical production of target protein using antisense RNA expression vectors

Cited by 8 publications

References 19 publications

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Oligonucleotide Squelching Reveals the Mechanism of Estrogen Receptor Autologous Down-Regulation

Comparison of three 3′ non-coding regions in Schizosaccharomyces pombe expression vectors: efficiencies of transcription termination and mRNA 3′-end formation

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