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来自鱼腥藻7120营养细胞的一种序列特异性DNA结合因子（VF1）与xisA上游区域的三个相邻位点结合。

A sequence-specific DNA-binding factor (VF1) from Anabaena sp. strain PCC 7120 vegetative cells binds to three adjacent sites in the xisA upstream region.

作者信息

Chastain C J, Brusca J S, Ramasubramanian T S, Wei T F, Golden J W

机构信息

Department of Biology, Texas A&M University, College Station 77843.

出版信息

J Bacteriol. 1990 Sep;172(9):5044-51. doi: 10.1128/jb.172.9.5044-5051.1990.

DOI:10.1128/jb.172.9.5044-5051.1990
PMID:2118506
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC213161/
Abstract

A DNA-binding factor (VF1) partially purified from Anabaena sp. strain PCC 7120 vegetative cell extracts by heparin-Sepharose chromatography was found to have affinity for the xisA upstream region. The xisA gene is required for excision of an 11-kilobase element from the nifD gene during heterocyst differentiation. Previous studies of the xisA upstream sequences demonstrated that deletion of this region is required for the expression of xisA from heterologous promoters in vegetative cells. Mobility shift assays with a labeled 250-base-pair fragment containing the binding sites revealed three distinct DNA-protein complexes. Competition experiments showed that VF1 also bound to the upstream sequences of the rbcL and glnA genes, but the rbcL and glnA fragments showed only single complexes in mobility shift assays. The upstream region of the nifH gene formed a weak complex with VF1. DNase footprinting and deletion analysis of the xisA binding site mapped the binding to a 66-base-pair region containing three repeats of the consensus recognition sequence ACATT.

摘要

通过肝素-琼脂糖层析从鱼腥藻属PCC 7120营养细胞提取物中部分纯化得到的一种DNA结合因子（VF1），被发现对xisA上游区域具有亲和力。xisA基因是异型胞分化过程中从nifD基因切除一个11千碱基元件所必需的。先前对xisA上游序列的研究表明，该区域的缺失是营养细胞中异源启动子表达xisA所必需的。用包含结合位点的标记250碱基对片段进行的迁移率变动分析揭示了三种不同的DNA-蛋白质复合物。竞争实验表明，VF1也与rbcL和glnA基因的上游序列结合，但rbcL和glnA片段在迁移率变动分析中仅显示单一复合物。nifH基因的上游区域与VF1形成弱复合物。对xisA结合位点的DNase足迹分析和缺失分析将结合定位到一个66碱基对区域，该区域包含共有识别序列ACATT的三个重复。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/5165bda464b7/jbacter00123-0320-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/4995c41d2378/jbacter00123-0316-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/df38a8daa900/jbacter00123-0317-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/db96480108fe/jbacter00123-0318-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/f05edd6a0478/jbacter00123-0319-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/5165bda464b7/jbacter00123-0320-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/4995c41d2378/jbacter00123-0316-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/df38a8daa900/jbacter00123-0317-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/db96480108fe/jbacter00123-0318-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/f05edd6a0478/jbacter00123-0319-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/045f/213161/5165bda464b7/jbacter00123-0320-a.jpg

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