Characterization of Helicobacter pylori lytic transglycosylases Slt and MltD.
Peptidoglycan (PG) is a cell wall heteropolymer that is essential for cell integrity. PG hydrolases participate in correct assembly of the PG layer and have been shown to be required for cell division, cell daughter separation, and maintenance of bacterial morphology. In silico analysis of the Helicobacter pylori genome resulted in identification of three potential hydrolases, Slt, MltD, and AmiA. This study was aimed at determining the roles of the putative lytic transglycosylases, Slt and MltD, in H. pylori morphology, growth, and PG metabolism. Strain 26695 single mutants were constructed using a nonpolar kanamycin cassette. The slt and mltD mutants formed normal bacillary and coccoid bacteria in the exponential and stationary phases, respectively. The slt and mltD mutants had growth rates comparable to the growth rate of the parental strain. However, the mltD mutant exhibited enhanced survival in the stationary phase compared to the wild type or the slt mutant. PG was purified from exponentially growing bacteria and from bacteria in the stationary phase, and its muropeptide composition was analyzed by high-pressure liquid chromatography. This analysis revealed changes in the muropeptide composition indicating that MltD and Slt have lytic transglycosylase activities. Glycan strand analysis suggested that Slt and MltD have exo and endo types of lytic transglycosylase activity, indicating that Slt is involved mainly in PG turnover and MltD is involved mainly in rearrangement of the PG layer. In this study, we determined the distinct roles of the lytic transglycosylases Slt and MltD in PG metabolism.
Chaput
Catherine
C
Unité de Pathogénie Bactérienne des Muqueuses, Department of Microbiology, Institut Pasteur, 28 Rue du Dr. Roux, 75724 Paris cedex 15, France.
Labigne
Agnès
A
Boneca
Ivo G
IG
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000201:enzymology / Q000648:ultrastructure; D023061:Gene Order; D016680:Genome, Bacterial; D016480:Helicobacter pylori / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D007527:Isoenzymes; D010457:Peptidoglycan; D009238:N-Acetylmuramoyl-L-alanine Amidase