A trigger enzyme in Mycoplasma pneumoniae: impact of the glycerophosphodiesterase GlpQ on virulence and gene expression

• Published in: PLoS pathogens Vol. 7; no. 9; p. e1002263
• Main Authors: Schmidl, Sebastian R, Otto, Andreas, Lluch-Senar, Maria, Piñol, Jaume, Busse, Julia, Becher, Dörte, Stülke, Jörg
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.09.2011; Public Library of Science (PLoS)
• Subjects: Bacteriology; Biology; Cell Survival - drug effects; Enzymes; Gene expression; Gene Expression Regulation, Bacterial; Genetic aspects; Glycerolphosphate Dehydrogenase - metabolism; Health aspects; HeLa Cells; Humans; Hydrogen Peroxide - metabolism; Hydrogen Peroxide - pharmacology; Microbiology; Mycoplasma pneumoniae; Mycoplasma pneumoniae - genetics; Phosphodiesterases; Phosphoric Diester Hydrolases - genetics; Phosphoric Diester Hydrolases - physiology; Physiological aspects; Proteins; Proteome; Virulence - genetics; Germany; Spain
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1002263

Mycoplasma pneumoniae is a causative agent of atypical pneumonia. The formation of hydrogen peroxide, a product of glycerol metabolism, is essential for host cell cytotoxicity. Phosphatidylcholine is the major carbon source available on lung epithelia, and its utilization requires the cleavage of deacylated phospholipids to glycerol-3-phosphate and choline. M. pneumoniae possesses two potential glycerophosphodiesterases, MPN420 (GlpQ) and MPN566. In this work, the function of these proteins was analyzed by biochemical, genetic, and physiological studies. The results indicate that only GlpQ is an active glycerophosphodiesterase. MPN566 has no enzymatic activity as glycerophosphodiesterase and the inactivation of the gene did not result in any detectable phenotype. Inactivation of the glpQ gene resulted in reduced growth in medium with glucose as the carbon source, in loss of hydrogen peroxide production when phosphatidylcholine was present, and in a complete loss of cytotoxicity towards HeLa cells. All these phenotypes were reverted upon complementation of the mutant. Moreover, the glpQ mutant strain exhibited a reduced gliding velocity. A comparison of the proteomes of the wild type strain and the glpQ mutant revealed that this enzyme is also implicated in the control of gene expression. Several proteins were present in higher or lower amounts in the mutant. This apparent regulation by GlpQ is exerted at the level of transcription as determined by mRNA slot blot analyses. All genes subject to GlpQ-dependent control have a conserved potential cis-acting element upstream of the coding region. This element overlaps the promoter in the case of the genes that are repressed in a GlpQ-dependent manner and it is located upstream of the promoter for GlpQ-activated genes. We may suggest that GlpQ acts as a trigger enzyme that measures the availability of its product glycerol-3-phosphate and uses this information to differentially control gene expression.