Sequence analysis, characterization and CO-specific transcription of the cox gene cluster on the megaplasmid pHCG3 of Oligotropha carboxidovorans.

Abstract

Sequence, transcriptional, mutational and physiological analyses indicate that the carbon monoxide (CO) dehydrogenase of Oligotropha carboxidovorans is an integral and unique part of an elaborate CO oxidizing system. It is encoded by the 14.5kb gene cluster coxBCMSLDEFGHIK residing on the 128kb megaplasmid pHCG3. The CO dehydrogenase structural genes coxMSL are flanked by nine accessory genes arranged as the cox gene cluster. The cox genes are specifically and coordinately transcribed under chemolithoautotrophic conditions in the presence of CO as carbon and energy source. With the exception of CoxB and CoxK, all deduced products of the cox genes of O. carboxidovorans have counterparts in so far uncharacterized gene clusters of Pseudomonas thermocarboxydovorans, Hydrogenophaga pseudoflava, Bradyrhizobium japonicum, and Mycobacterium tuberculosis. Transposon mutagenesis suggests a function of CoxH and CoxI in the interaction of CO dehydrogenase with the cytoplasmic membrane. The specific functions of the other accessory Cox proteins are difficult to envisage right now, as the polypeptides do not show significant homologies with functionally characterized proteins in the databases. In addition to the clustered cox genes, mutational analyses have identified the genes lon, cycH and orfX which reside on the plasmid pHCG3. The Lon protease, the CycH protein and the unknown orfX gene product have essential functions in the utilization of CO.