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通过酵母DNA结合蛋白的功能分析揭示转录激活因子、沉默子和端粒之间的联系

Connections between transcriptional activators, silencers, and telomeres as revealed by functional analysis of a yeast DNA-binding protein.

作者信息

Buchman A R, Lue N F, Kornberg R D

机构信息

Department of Cell Biology, Stanford University School of Medicine, California 94305.

出版信息

Mol Cell Biol. 1988 Dec;8(12):5086-99. doi: 10.1128/mcb.8.12.5086-5099.1988.

DOI:10.1128/mcb.8.12.5086-5099.1988
PMID:3072472
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC365610/
Abstract

General regulatory factor I (GRFI) is a yeast protein that binds in vitro to specific DNA sequences at diverse genetic elements. A strategy was pursued to test whether GRFI functions in vivo at the sequences bound by the factor in vitro. Matches to a consensus sequence for GRFI binding were found in a variety of locations: upstream activating sequences (UASs), silencers, telomeres, and transcribed regions. All occurrences of the consensus sequence bound both crude and purified GRFI in vitro. All binding sites for GRFI, regardless of origin, provided UAS function in test plasmids. Also, GRFI binding sites specifically stimulated transcription in a yeast in vitro system, indicating that GRFI can function as a positive transcription factor. The stimulatory effect of GRFI binding sites at UASs for the PYK1 and ENO1 genes is significantly enhanced by flanking DNA elements. By contrast, regulatory sequences that flank the GRFI binding site at HMR E convert this region to a transcriptional silencer.

摘要

通用调控因子I（GRFI）是一种酵母蛋白，它在体外可与不同遗传元件上的特定DNA序列结合。我们采用了一种策略来测试GRFI在体内是否在其体外结合的序列上发挥作用。在多种位置发现了与GRFI结合的共有序列的匹配：上游激活序列（UAS）、沉默子、端粒和转录区域。共有序列的所有出现情况在体外均能结合粗提物和纯化的GRFI。GRFI的所有结合位点，无论其来源如何，在测试质粒中都具有UAS功能。此外，GRFI结合位点在酵母体外系统中能特异性地刺激转录，表明GRFI可作为一种正转录因子发挥作用。PYK1和ENO1基因UAS处的GRFI结合位点的刺激作用会因侧翼DNA元件而显著增强。相比之下，HMR E处GRFI结合位点侧翼的调控序列会将该区域转变为转录沉默子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/601393b1d3c2/molcellb00072-0049-a.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/d5ccecc8437f/molcellb00072-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/8879b0bb4996/molcellb00072-0048-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/601393b1d3c2/molcellb00072-0049-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/080c39f2456c/molcellb00072-0043-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/6d5013bfcedf/molcellb00072-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/bd7d221f38e1/molcellb00072-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/aa7d98de904a/molcellb00072-0045-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/d5ccecc8437f/molcellb00072-0046-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/8879b0bb4996/molcellb00072-0048-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/09fd/365610/601393b1d3c2/molcellb00072-0049-a.jpg

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