The role of the genes nrf EFG and ccmFH in cytochrome c biosynthesis in Escherichia coli.

It has been suggested that two groups of Escherichia coli genes, the ccm genes located in the 47-min region and the nrfEFG genes in the 92-min region of the chromosome, are involved in cytochrome c biosynthesis during anaerobic growth. The involvement of the products of these genes in cytochrome c synthesis, assembly and secretion has now been investigated. Despite their similarity to other bacterial cytochrome c assembly proteins, NrfE, F and G were found not to be required for the biosynthesis of any of the c-type cytochromes in E. coli. Furthermore, these proteins were not required for the secretion of the periplasmic cytochromes, cytochrome C550 and cytochrome C552, or for the correct targeting of the NapC and NrfB cytochromes to the cytoplasmic membrane. NrfE and NrfG are required for formate-dependent nitrite reduction (the Nrf pathway), which involves at least two c-type cytochromes, cytochrome C552 and NrfB, but NrfF is not essential for this pathway. Genes similar to nrfE, nrfF and nrfG are present in the E. coli nap-ccm locus at minute 47. CcmF is similar to NrfE, the N-terminal region of CcmH is similar to NrfF and the C-terminal portion of CcmH is similar to NrfG. In contrast to NrfF, the N-terminal, NrfF-like portion of CcmH is essential for the synthesis of all c-type cytochromes. Conversely, the NrfG-like C-terminal region of CcmH is not essential for cytochrome c biosynthesis. The data are consistent with proposals from this and other laboratories that CcmF and CcmH form part of a haem lyase complex required to attach haem c to C-X-X-C-H haem-binding domains. In contrast, NrfE and NrfG are proposed to fulfill a more specialised role in the assembly of the formate-dependent nitrite reductase.