Catalase-peroxidases of Legionella pneumophila: cloning of the katA gene and studies of KatA function.
Legionella pneumophila, the causative organism of Legionnaires' pneumonia, contains two enzymes with catalatic and peroxidatic activity, KatA and KatB. To address the issue of redundant, overlapping, or discrete in vivo functions of highly homologous catalase-peroxidases, the gene for katA was cloned and its function was studied in L. pneumophila and Escherichia coli and compared with prior studies of katB in this laboratory. katA is induced during exponential growth and is the predominant peroxidase in stationary phase. When katA is inactivated, L. pneumophila is more sensitive to exogenous hydrogen peroxide and less virulent in the THP-1 macrophage cell line, similar to katB. Catalatic-peroxidatic activity with different peroxidatic cosubstrates is comparable for KatA and KatB, but KatA is five times more active towards dianisidine. In contrast with these examples of redundant or overlapping function, stationary-phase survival is decreased by 100- to 10,000-fold when katA is inactivated, while no change from wild type is seen for the katB null. The principal clue for understanding this discrete in vivo function was the demonstration that KatA is periplasmic and KatB is cytosolic. This stationary-phase phenotype suggests that targets sensitive to hydrogen peroxide are present outside the cytosol in stationary phase or that the peroxidatic activity of KatA is critical for stationary-phase redox reactions in the periplasm, perhaps disulfide bond formation. Since starvation-induced stationary phase is a prerequisite to acquisition of virulence by L. pneumophila, further studies on the function and regulation of katA in stationary phase may give insights on the mechanisms of infectivity of this pathogen.
Bandyopadhyay
P
P
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Steinman
H M
HM
D000332:Aerobiosis; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D007424:Intracellular Fluid / Q000378:metabolism; D016952:Legionella pneumophila / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D008264:Macrophages / Q000166:cytology / Q000382:microbiology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D013379:Substrate Specificity
D004269:DNA, Bacterial; D006861:Hydrogen Peroxide