Localization and expression of the Bacillus subtilis dl-endopeptidase LytF are influenced by mutations in LTA synthases and glycolipid anchor synthetic enzymes

Kiriyama, Yuuka; Yazawa, Kazuya; Tanaka, Tatsuhito; Yoshikawa, Ritsuko; Yamane, Hisaya; Hashimoto, Masayuki; Sekiguchi, Junichi; Yamamoto, Hiroki

doi:10.1099/mic.0.080366-0

Cited by 10 publications

(20 citation statements)

References 50 publications

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“…In addition to these phenotypes, a quadruple ltaS homolog mutant shows a loss of LTA (7), an aberrant twisted morphology, slower growth (6), and reduced adsorption of rare earth elements (8,9). Moreover, we reported that D -dependent transcription of lytF, which encodes a DL-endopeptidase that functions in cell separation, is reduced in an ltaS mutant and is nearly absent in multiple mutants of ltaS and its homologs (10). Interestingly, we also found that lytF transcription was repressed in tagO-null mutant cells, which lack WTA.…”

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confidence: 88%

“…The sample preparation for immunofluorescence microscopy (IFM) was described previously (10,23) and used with minor modifications, as follows. An anti-DYKDDDDK tag monoclonal antibody (1:400; Wako Pure Chemical Industries) was used as the primary antibody, and an Alexa Fluor 555 F(ab=) 2 fragment of goat anti-mouse IgG (HϩL) (1:1,200; Life Technologies) was used as the secondary antibody.…”

Section: Methodsmentioning

confidence: 99%

“…After electrophoresis, Western blot analysis of the LytE-6ϫFLAG fusion protein was performed as described previously (23). An anti-DYKDDDDK tag monoclonal antibody (Wako Pure Chemical Industries) was used as the primary antibody, and an anti-mouse IgG, horseradish peroxidase (HRP)-linked whole antibody (GE Healthcare) was used as the secondary antibody (both at 1:10,000 for FLAGspecific detection as described previously) (10). For the Western blot analysis of LTA, the sample preparation, SDS-PAGE, and blotting were performed as described previously (7).…”

Section: Methodsmentioning

confidence: 99%

“…Previous reports have shown that cells lacking LTA grow slowly and have an aberrant twisted chained morphology (6,10). In addition, when WTA was abolished, cells grew slowly, showed a swelled morphology, and formed clumps (20).…”

mentioning

confidence: 99%

“…These results indicate that LTA and WTA, which are similar anionic polymers with a poly(GroP) backbone located on the B. subtilis cell surface, are required for temporal D -dependent lytF expression. Moreover, both WTA and LTA hinder LytF localization in the cylindrical part of the cell (10,11). In addition to septum PG digestion by LytF and CwlS, sidewall PG hydrolysis, which is required for nascent PG incorporation, is catalyzed by LytE and CwlO during vegetative growth (12,13).…”

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confidence: 99%

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Teichoic Acid Polymers Affect Expression and Localization of dl -Endopeptidase LytE Required for Lateral Cell Wall Hydrolysis in Bacillus subtilis

et al. 2016

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In Bacillus subtilis, the DL-endopeptidase LytE is responsible for lateral peptidoglycan hydrolysis during cell elongation. We found that I -dependent transcription of lytE is considerably enhanced in a strain with a mutation in ltaS, which encodes a major lipoteichoic acid (LTA) synthase. Similar enhancements were observed in mutants that affect the glycolipid anchor and wall teichoic acid (WTA) synthetic pathways. Immunofluorescence microscopy revealed that the LytE foci were considerably increased in these mutants. The localization patterns of LytE on the sidewalls appeared to be helix-like in LTA-defective or WTAreduced cells and evenly distributed on WTA-depleted or -defective cell surfaces. These results strongly suggested that LTA and WTA affect both I -dependent expression and localization of LytE. Interestingly, increased LytE localization along the sidewall in the ltaS mutant largely occurred in an MreBH-independent manner. Moreover, we found that cell surface decorations with LTA and WTA are gradually reduced at increased culture temperatures and that LTA rather than WTA on the cell surface is reduced at high temperatures. In contrast, the amount of LytE on the cell surface gradually increased under heat stress conditions. Taken together, these results indicated that reductions in these anionic polymers at high temperatures might give rise to increases in SigI-dependent expression and cell surface localization of LytE at high temperatures. IMPORTANCEThe bacterial cell wall is required for maintaining cell shape and bearing environmental stresses. The Gram-positive cell wall consists of mesh-like peptidoglycan and covalently linked wall teichoic acid and lipoteichoic acid polymers. It is important to determine if these anionic polymers are required for proliferation and environmental adaptation. Here, we demonstrated that these polymers affect the expression and localization of a peptidoglycan hydrolase LytE required for lateral cell wall elongation. Moreover, we found that cell surface decorations with teichoic acid polymers are substantially decreased at high temperatures and that the peptidoglycan hydrolase is consequently increased. These findings suggest that teichoic acid polymers control lateral peptidoglycan hydrolysis by LytE, and bacteria drastically change their cell wall content to adapt to their environment.

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mentioning

confidence: 88%

Section: Methodsmentioning

confidence: 99%