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Bacillus subtilis cytochromec oxidasecaa 3 is encoded by thectaCDEF genes at thectaABCDEF locus, with thectaBCDEF genes organized as an operon-like unit. A dyad symmetry sequence and a catabolite response element homolog can be recognized in the 240-bp intercistronic region betweenctaB andctaC. ctaB′-lacZ andctaBCD′-lacZ transcriptional fusions integrated at the native locus were used to study catabolite effects on transcription of thectaB andctaCDEF genes. In Schaeffer’s medium lacking glucose,ctaBCD′-lacZ was expressed at a very low level during the exponential phase, and expression increased about 30-fold 2 h after entry into the stationary phase. In the presence of 0.5% glucose,ctaBCD′-lacZ expression was totally repressed. In contrast toctaBCD′-lacZ ,ctaB′-lacZ was constitutively expressed regardless of carbon source. ThectaCDEF genes were separated fromctaB by insertion of plasmids carrying selectable markers in such a way that thectaCDEF andctaB transcription units remained intact. Enzymatic assays ofcaa 3 with these constructs, showed thatctaCDEF was not expressed independently ofctaB . Also, when a ′ctaB-ctaC′-lacZ fusion (containing thectaB-ctaC intercistronic region) was placed at a remote nonessential locus, β-galactosidase activity could not be detected. The absence of a promoter in thectaB-ctaC intercistronic space also was indicated by the inability to detectctaC -specific transcripts with RNase protection assays, primer extension, and rapid amplification of 5′ cDNA ends. Direct mRNA measurements showed that, in the presence of 0.5% glucose,ctaBCDEF transcripts terminated at the 3′ end of the putative stem-loop structure and the distal portion was down-regulated. A possible mechanism forctaCDEF gene regulation is suggested. Catabolite repression ofctaBCD′-lacZ was partly dependent on CcpA but was independent of HPr. The expression ofctaBCDEF also appears to require thestrC ,ctaA , andresD-resE gene products.

Catabolite Regulation of the Bacillus subtilis ctaBCDEF Gene Cluster