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Effect of the Nuclear Factors EmBP1 and Viviparousl on the Transcription of the Em Gene in HeLa Nuclear Extracts
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BibTeX
@MISC{Razik_effectof,
author = {Mona A. Razik and Ralph S. Quatrano},
title = {Effect of the Nuclear Factors EmBP1 and Viviparousl on the Transcription of the Em Gene in HeLa Nuclear Extracts},
year = {}
}
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Abstract
Templates constructed from the wheat Em and maize rab28 promoters are efficiently and accurately transcribed in the well-characterized cell-free transcription system prepared from HeLa nuclei. Deletion analysis of the Em promoter indicates that a G-box (CACGTG) element (Em1 b) is required for transcription. USF, a Myc transcription factor in HeLa nuclear extracts, activates transcription by binding to Emlb, as shown by the ability of an antibody raised against USF to inhibit transcription and to interfere with Em1 b complex formation in an electrophoretic mobility shift assay. The addition of the recombinant Viviparousl protein from maize to HeLa nuclear extracts specifically stimulated transcription of the Em promoter but was dependent on the presence of USF in the extract. In USF-depleted extracts, the addition of recombinant EmBP1, a basic leucine zipper transcription factor from wheat, activated transcription through Em1 b as well as from a similar G-box in the adenovirus major late promoter. Our study demonstrates that the basic transcriptional apparatus in HeLa nuclear extract supports transcription from plant promoters and can be used to assay the function of certain plant nuclear proteins, thereby helping to determine their effects on transcription.
Keyphrases
hela nuclear extract nuclear factor embp1 em gene em promoter adenovirus major late promoter activates transcription certain plant nuclear protein electrophoretic mobility shift assay hela nuclear extract support transcription basic leucine zipper transcription factor rab28 promoter em1 complex formation wheat em recombinant embp1 usf-depleted extract well-characterized cell-free transcription system plant promoter deletion analysis myc transcription factor similar g-box hela nucleus basic transcriptional apparatus recombinant viviparousl protein
