Spacer mutations in the lac ps promoter

Abstract

Mutations have been constructed that delete either one or two base pairs near position -19 in the lac ps promoter. Deletion of either of two adjacent base pairs increases the rates of open complex formation by nearly on order of magnitude. Two promoters that have different single-base deletions are indistinguishable by either their rates of open complex formation or stability of the open complexes once formed. However, simultaneous deletion of both base pairs produces a promoter that forms complexes at a rate similar to that of the unmodified DNA sequence. The maximal rate of open complex formation is achieved at a spacer length of 17 base pairs, the most frequently occurring spacer length among promoters. These results suggest that the spacing between the two strongly conserved regions of sequence homology is an important determinant of the rate of open complex formation. A model is suggested that proposes that three important promoter elements, the -10 region, the -35 region, and the spacer region, act simultaneously to facilitate open complex formation by RNA polymerase.