CONSTRUCTION AND FUNCTION OF CHIMERIC BETA-SUBUNITS CONTAINING REGIONS FROM THE BETA-SUBUNITS OF THE F1F0 ATPASES OF ESCHERICHIA-COLI AND BACILLUS-MEGATERIUM

The highly conserved beta subunit of the Escherichia coli F1F0 ATPase was divided into three sections, each of which was exchanged with the homologous section of the beta subunit of the obligate aerobe Bacillus megaterium. Plasmids coding for the resultant six chimeric beta subunits varied in their abilities to complement two E. colt beta mutants as measured by testing transformed cells for aerobic growth on a nonfermentable carbon source or anaerobic growth on rich medium containing glucose. Two chimeras were able to restore both growth on succinate and anaerobic growth on rich medium. The genetic results corresponded to increased levels of membrane-bound ATPase and ATP synthase activities. These chimeric subunits were therefore capable of being assembled into functional E. coli ATPase complexes. The results indicate that chimeric beta subunits can be used to analyze assembly of the beta subunit and that the final 181 amino acids of the beta subunit might contain a region involved in functional energy coupling.