Sequence and properties of operator mutations in the bio operon of Escherichia coli

Abstract

The nucleotide-sequence changes occurring in newly isolated operator-constitutive mutations of the divergently transcribed bio operon have been determined. The observed point mutations are single GC leads to AT changes which occur at two symmetrical points in the hyphenated inverted repeat present in the control region. The changes at position -15 (with respect to the center of the inverted repeat) cause constitutivity of leftward operon expression and decreased expression of bioB, due to alteration of the -35 region of the rightward promoter. The change at position +15 is identical to one of the changes that Otsuka and Abelson (1978) detected in the bio p98 bio o34 double mutant. The location of the bio p131::IS1 insertion, which affects both leftward and rightward transcription, is also within the operator region. Both of the operator-constitutive mutations and the bio p131 insertion cause decreased repressor binding in vivo as shown by repressor titration tests on multicopy plasmids which bear them. The operator-constitutive mutations also decrease repressor binding in vitro, where added repressor fails to protect the TaqI site that is protected by repressor in bio o+ DNA. These results confirm several aspects of the model for bio operon regulation proposed by Otsuka and Abelson (1978).