title: Synthetic lethality of the lytE cwlO genotype in Bacillus subtilis is caused by lack of D,L-endopeptidase activity at the lateral cell wall.
abstract: Bacterial peptidoglycan acts as an exoskeleton to protect the bacterial cell. Although peptidoglycan biosynthesis by penicillin-binding proteins is well studied, few studies have described peptidoglycan disassembly, which is necessary for a dynamic structure that allows cell growth. In Bacillus subtilis, more than 35 genes encoding cell wall lytic enzymes have been identified; however, only two D,L-endopeptidases (lytE and cwlO) are involved in cell proliferation. In this study, we demonstrated that the D,L-endopeptidase activity at the lateral cell wall is essential for cell proliferation. Inactivation of LytE and CwlO by point mutation of the catalytic residues caused cell growth defects. However, the forced expression of LytF or CwlS, which are paralogs of LytE, did not suppress lytE cwlO synthetic lethality. Subcellular localization studies of these D,L-endopeptidases showed LytF and CwlS at the septa and poles, CwlO at the cylindrical part of the cell, and LytE at the septa and poles as well as the cylindrical part. Furthermore, construction of N-terminal and C-terminal domain-swapped enzymes of LytE, LytF, CwlS, and CwlO revealed that localization was dependent on the N-terminal domains. Only the chimeric proteins that were enzymatically active and localized to the sidewall were able to suppress the synthetic lethality, suggesting that the lack of D,L-endopeptidase activity at the cylindrical part of the cell leads to a growth defect. The functions of LytE and CwlO in cell morphogenesis were discussed.
authors.affiliation: International Young Researchers Empowerment Center, Shinshu University, Ueda, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Hashimoto
authors.forename: Seika
authors.initials: S
authors.lastname: Ooiwa
authors.forename: Junichi
authors.initials: J
authors.lastname: Sekiguchi
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001433:Bacteriolysis; D002473:Cell Wall / Q000737:chemistry / Q000201:enzymology*; D010450:Endopeptidases / Q000378:metabolism*; D005798:Genes, Bacterial; D005838:Genotype; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism; D017354:Point Mutation
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan; D010450:Endopeptidases; C112181:LytE protein, Bacillus subtilis; D009238:N-Acetylmuramoyl-L-alanine Amidase