Leisy D J, Rohrmann G F
Department of Microbiology, Oregon State University, Corvallis, Oregon 97331, USA.
Virology. 2000 Aug 15;274(1):196-202. doi: 10.1006/viro.2000.0452.
Missing contact footprinting with formic acid as a modifying reagent was used to examine specific IE-1 binding contacts to double-stranded oligonucleotides that contained either a consensus hr repeat sequence or a sequence from the pe38 promoter, which is down regulated by IE-1. The hr repeat sequences contain two consensus IE-1 binding motifs (IBMs) flanking a central EcoRI site that are oriented in opposite directions with respect to each other. IE-1 was found to contact regions including both IBMs. The bases footprinted in the top strand included the left IBM (IBM-A), whereas bases in the bottom strand were footprinted in a region that included IBM-B and part of IBM-A. When substitution mutations were introduced into either IBM, bases on both strands of the remaining IBM were strongly footprinted. As with the hr IBM-mutant constructs, bases footprinted in the pe38 promoter construct included both strands of the single IBM.
以甲酸作为修饰试剂的缺失接触足迹法,用于检测IE-1与双链寡核苷酸的特定结合接触,这些双链寡核苷酸包含一个共有hr重复序列或来自pe38启动子的序列,pe38启动子受IE-1下调。hr重复序列包含两个共有IE-1结合基序(IBM),位于一个中央EcoRI位点两侧,它们彼此方向相反。发现IE-1与包括两个IBM的区域接触。在上链中被足迹的碱基包括左侧IBM(IBM-A),而在下链中被足迹的碱基位于一个包括IBM-B和部分IBM-A的区域。当将取代突变引入任一IBM时,剩余IBM的两条链上的碱基都被强烈足迹。与hr IBM突变体构建体一样,在pe38启动子构建体中被足迹的碱基包括单个IBM的两条链。