Mutational analysis of the dimethylsulfoxide respiratory (dor) operon of Rhodobacter capsulatus.
Four genes, dorC, dorD, dorB and dorR of the DMSO respiratory gene cluster of Rhodobacter capsulatus have been identified and sequenced. dorC encodes a pentahaem c-type cytochrome of the NirT class and the derived DorC protein sequence shows highest similarity to TorC from the Escherichia coli trimethylamine-N-oxide (TMAO) respiratory system. Mutagenesis of dorC resulted in the loss of a 46 kDa haem-staining polypeptide from membranes of R. capsulatus. dorD encodes a protein with highest sequence similarity to TorD from the E. coli TMAO respiratory system. DMSO reductase polypeptide (DorA) could not be detected in cell-free extracts of a dorD mutant and it is suggested that DorD has a role in stabilizing the DorA apo-protein prior to insertion of the pterin molybdenum cofactor. dorB encodes a protein with highest sequence similarity to NapD of Paracoccus denitrificans. Mutagenesis of dorB reduced the activity of DMSO reductase and led to the accumulation of a larger form of the enzyme that is presumed to represent a cytoplasmic precursor polypeptide. It is suggested that DorB has a role in the biogenesis of DMSO reductase prior to its secretion into the periplasm. dorR is transcribed in the opposite direction to dorC. The derived amino acid sequence of DorR indicates that it is a response regulator and mutation of dorR shows that it is essential for expression of the dorCDA operon. Expression of a chromosomal dorA::lacZ fusion was also dependent on the transcriptional regulator Fnr. The intergenic region between dorR and dorC contains four putative binding sites for DorR but no binding site for Fnr was identified.
Shaw
Anthony L
AL
Department of Journal of General Microbiology, The University of Queensland, Brisbane 4072, Australia.
Leimkuler
Silke
S
Lehrstuhl für Biologie der Mikrooganismen, Ruhr Universitat Bochum, D-44780, Bochum, Germany.
Klipp
Werner
W
Lehrstuhl für Biologie der Mikrooganismen, Ruhr Universitat Bochum, D-44780, Bochum, Germany.
Hanson
Graeme R
GR
Centre for Magnetic Resonance, The University of Queensland, Brisbane 4072, Australia.
McEwan
Alastair G
AG
Department of Journal of General Microbiology, The University of Queensland, Brisbane 4072, Australia.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D003574:Cytochrome c Group / Q000096:biosynthesis / Q000235:genetics; D003580:Cytochromes / Q000096:biosynthesis; D004252:DNA Mutational Analysis; D004121:Dimethyl Sulfoxide / Q000378:metabolism*; D004790:Enzyme Induction; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D009876:Operon; D016354:Rhodobacter capsulatus / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D003580:Cytochromes; C120747:DorB protein, Rhodobacter capsulatus; C120749:DorC protein, Rhodobacter sphaeroides; C120748:DorR protein, Rhodobacter capsulatus; D014157:Transcription Factors; D004121:Dimethyl Sulfoxide