Analysis of regulatory sequence upstream of the E. coli uvrB gene; involvement of the DnaA protein

A region located upstream of the uvrB promoters P1 and P2 was found to cause high plasmid loss when cloned in multicopy vectors. Two sequence elements responsible for this phenomenon were identified by mapping of spontaneous mutations that restore plasmid maintenance: a sequence known to have in vitro promoter activity and a partially overlapping sequence that shows extensive homology to recognition sites for the DnaA protein. Accordingly alterations in the level of DnaA protein in vivo were found to affect the extent of plasmid loss. A possible role for interaction of the DnaA protein with the region of interest is discussed in relation to regulation of uvrB expression.