Journal
MICROBIOLOGY-SGM
Volume 164, Issue 4, Pages 670-684Publisher
MICROBIOLOGY SOC
DOI: 10.1099/mic.0.000625
Keywords
Bacillus subtilis; two-component system; WalRK; LytE
Categories
Funding
- Japanese Society for the Promotion of Science [23380052, 16H04903, 16K14892, 15K18676]
- MEXT [S1311017]
- Grants-in-Aid for Scientific Research [16H04903, 15K18676, 16K14892, 23380052] Funding Source: KAKEN
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WalRK is an essential two-component signal transduction system that plays a central role in coordinating cell wall synthesis and cell growth in Bacillus subtilis. However, the physiological role of WalRK and its essentiality for growth have not been elucidated. We investigated the behaviour of WalRK during heat stress and its essentiality for cell proliferation. We determined that the inactivation of the walHI genes which encode the negative modulator of WalK, resulted in growth defects and eventual cell lysis at high temperatures. Screening of suppressor mutations revealed that the inactivation of LytE, an DL-endopeptidase, restored the growth of the Delta walHI mutant at high temperatures. Suppressor mutations that reduced heat induction arising from the walRK regulon were also mapped to the walK ORF. Therefore, we hypothesized that overactivation of LytE affects the phenotype of the Delta walHI mutant. This hypothesis was corroborated by the overexpression of the negative regulator of LytE, IseA and PdaC, which rescued the growth of the Delta walHI mutant at high temperatures. Elucidating the cause of the temperature sensitivity of the Delta walHI mutant could explain the essentiality of WalRK. We proved that the constitutive expression of lytE or cwlO using a synthetic promoter uncouples these expressions from WalRK, and renders WalRK nonessential in the pdaC and iseA mutant backgrounds. We propose that the essentiality of WalRK is derived from the coordination of cell wall metabolism with cell growth by regulating DL-endopeptidase activity under various growth conditions.
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