The Autographa californica nucleopolyhedrovirus IE-1 protein complex has two modes of specific DNA binding.
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.
Department of Microbiology, Oregon State University, Corvallis, Oregon 97331, USA. leisyd@bcc.orst.edu
D J
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Leisy
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Rohrmann
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D004279:DNA, Viral; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology; D016296:Mutagenesis; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism
C069183:Ac-IE-1 protein, Autographa californica nucleopolyhedrovirus; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D015534:Trans-Activators