Involvement of a membrane-bound class III adenylate cyclase in regulation of anaerobic respiration in Shewanella oneidensis MR-1.
Unlike other bacteria that use FNR to regulate anaerobic respiration, Shewanella oneidensis MR-1 uses the cyclic AMP receptor protein (CRP) for this purpose. Three putative genes, cyaA, cyaB, and cyaC, predicted to encode class I, class IV, and class III adenylate cyclases, respectively, have been identified in the genome sequence of this bacterium. Functional validation through complementation of an Escherichia coli cya mutant confirmed that these genes encode proteins with adenylate cyclase activities. Chromosomal deletion of either cyaA or cyaB did not affect anaerobic respiration with fumarate, dimethyl sulfoxide (DMSO), or Fe(III), whereas deletion of cyaC caused deficiencies in respiration with DMSO and Fe(III) and, to a lesser extent, with fumarate. A phenotype similar to that of a crp mutant, which lacks the ability to grow anaerobically with DMSO, fumarate, and Fe(III), was obtained when both cyaA and cyaC were deleted. Microarray analysis of gene expression in the crp and cyaC mutants revealed the involvement of both genes in the regulation of key respiratory pathways, such as DMSO, fumarate, and Fe(III) reduction. Additionally, several genes associated with plasmid replication, flagellum biosynthesis, and electron transport were differentially expressed in the cyaC mutant but not in the crp mutant. Our results indicated that CyaC plays a major role in regulating anaerobic respiration and may contribute to additional signaling pathways independent of CRP.
Charania
M A
MA
Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53211, USA.
Brockman
K L
KL
Zhang
Y
Y
Banerjee
A
A
Pinchuk
G E
GE
Fredrickson
J K
JK
Beliaev
A S
AS
Saffarini
D A
DA
D000262:Adenylyl Cyclases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D015153:Blotting, Western; D019069:Cell Respiration / Q000235:genetics / Q000502:physiology*; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D017384:Sequence Deletion; D020592:Shewanella / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology*
D001426:Bacterial Proteins; D000262:Adenylyl Cyclases