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

Microbiology ◽  
2014 ◽  
Vol 160 (12) ◽  
pp. 2639-2649 ◽  
Author(s):  
Yuuka Kiriyama ◽  
Kazuya Yazawa ◽  
Tatsuhito Tanaka ◽  
Ritsuko Yoshikawa ◽  
Hisaya Yamane ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Cell Separation ◽  
Vegetative Cell ◽  
Transcriptional Activity ◽  
Immunofluorescence Microscopy ◽  
Lipoteichoic Acid ◽  
Synthetic Enzymes ◽  
Principal Role ◽  
Artificial Induction ◽  
Glycolipid Anchor

Bacillus subtilis LytF plays a principal role in cell separation through its localization at the septa and poles on the vegetative cell surface. In this study, we found that a mutation in a major lipoteichoic acid (LTA) synthase gene – ltaS – results in a considerable reduction in the σD-dependent transcription of lytF. The lytF transcription was also reduced in mutants that affected glycolipid anchor biosynthesis. Immunofluorescence microscopy revealed that both the numbers of cells expressing LytF and the LytF foci in these mutants were decreased. In addition, the transcriptional activity of lytF was almost abolished in the double (ltaS yfnI), triple (ltaS yfnI yqgS), and quadruple (ltaS yfnI yqgS yvgJ) mutants during vegetative growth. Cell separation defects in these mutants were partially restored with artificial expression of LytF. Interestingly, when lytF transcription was induced in the ltaS single or multiple mutants, LytF was localized not only at the septum, but also along the sidewall. The amounts of LytF bound to cell wall in the single (ltaS) and double (ltaS yfnI) mutants gradually increased as compared with that in the WT strain, and those in the triple (ltaS yfnI yqgS) and quadruple mutants were almost similar to that in the double mutant. Moreover, reduction of the lytF transcription and chained cell morphology in the ltaS mutant were completely restored with artificial induction of the yqgS gene. These results strongly suggest that LTA influences the temporal, σD-dependent transcription of lytF and is an additional inhibitory component to the vegetative cell separation enzyme LytF.

scholarly journals Localization of the Vegetative Cell Wall Hydrolases LytC, LytE, and LytF on the Bacillus subtilis Cell Surface and Stability of These Enzymes to Cell Wall-Bound or Extracellular Proteases

2003 ◽  
Vol 185 (22) ◽  
pp. 6666-6677 ◽  
Author(s):  
Hiroki Yamamoto ◽  
Shin-ichirou Kurosawa ◽  
Junichi Sekiguchi
Keyword(s):  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Surface ◽  
Fusion Protein ◽  
Growth Phase ◽  
Vegetative Growth ◽  
Cell Separation ◽  
Vegetative Cell ◽  
Exponential Growth Phase ◽  
Cell Wall Hydrolases

ABSTRACT LytF, LytE, and LytC are vegetative cell wall hydrolases in Bacillus subtilis. Immunofluorescence microscopy showed that an epitope-tagged LytF fusion protein (LytF-3xFLAG) in the wild-type background strain was localized at cell separation sites and one of the cell poles of rod-shaped cells during vegetative growth. However, in a mutant lacking both the cell surface protease WprA and the extracellular protease Epr, the fusion protein was observed at both cell poles in addition to cell separation sites. This suggests that LytF is potentially localized at cell separation sites and both cell poles during vegetative growth and that WprA and Epr are involved in LytF degradation. The localization pattern of LytE-3xFLAG was very similar to that of LytF-3xFLAG during vegetative growth. However, especially in the early vegetative growth phase, there was a remarkable difference between the shape of cells expressing LytE-3xFLAG and the shape of cells expressing LytF-3xFLAG. In the case of LytF-3xFLAG, it seemed that the signals in normal rod-shaped cells were stronger than those in long-chain cells. In contrast, the reverse was found in the case of LytE-3xFLAG. This difference may reflect the dependence on different sigma factors for gene expression. The results support and extend the previous finding that LytF and LytE are cell-separating enzymes. On the other hand, we observed that cells producing LytC-3xFLAG are uniformly coated with the fusion protein after the middle of the exponential growth phase, which supports the suggestion that LytC is a major autolysin that is not associated with cell separation.

scholarly journals Alanylated lipoteichoic acid primer in Bacillus subtilis

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 155 ◽  
Author(s):  
Yu Luo
Keyword(s):  
Bacillus Subtilis ◽  
Lipoteichoic Acid ◽  
Head Group ◽  
Host Immune System ◽  
Type I ◽  
Bacterial Survival ◽  
Gram Positive Bacteria ◽  
Type Iv ◽  
Glycolipid Anchor ◽  
Group D

Lipoteichoic acid is a major lipid-anchored polymer in Gram-positive bacteria such asBacillus subtilis. This polymer typically consists of repeating phosphate-containing units and therefore has a predominant negative charge. The repeating units are attached to a glycolipid anchor which has a diacylglycerol (DAG) moiety attached to a dihexopyranose head group. D-alanylation is known as the major modification of type I and type IV lipoteichoic acids, which partially neutralizes the polymer and plays important roles in bacterial survival and resistance to the host immune system. The biosynthesis pathways of the glycolipid anchor and lipoteichoic acid have been fully characterized. However, the exact mechanism of D-alanyl transfer from the cytosol to cell surface lipoteichoic acid remains unclear. Here I report the use of mass spectrometry in the identification of possible intermediate species in the biosynthesis and D-alanylation of lipoteichoic acid: the glycolipid anchor, nascent lipoteichoic acid primer with one phosphoglycerol unit, as well as mono- and di-alanylated forms of the lipoteichoic acid primer. Monitoring these species as well as the recently reported D-alanyl-phosphatidyl glycerol should aid in shedding light on the mechanism of the D-alanylation pathway of lipoteichoic acid.

scholarly journals Alanylated lipoteichoic acid primer in Bacillus subtilis

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 155 ◽  
Author(s):  
Yu Luo
Keyword(s):  
Bacillus Subtilis ◽  
Negative Charge ◽  
Lipoteichoic Acid ◽  
Head Group ◽  
Host Immune System ◽  
Bacterial Survival ◽  
Intermediate Species ◽  
Gram Positive Bacteria ◽  
Glycolipid Anchor ◽  
Group D

Lipoteichoic acid is a major lipid-anchored polymer in Gram-positive bacteria such asBacillus subtilis. This polymer typically consists of repeating phosphoglycerol or phosphoribitol units and therefore has a predominant negative charge. The repeating units are attached to a glycolipid anchor which has a diacylglycerol (DAG) moiety attached to a dihexopyranose head group. D-alanylation is known as the major modification of lipoteichoic acid, which partially neutralizes the polymer and plays important roles in bacterial survival and resistance to the host immune system. The biosynthesis pathways of the glycolipid anchor and lipoteichoic acid have been fully characterized. However, the exact mechanism of D-alanyl transfer from the cytosol to cell surface lipoteichoic acid remains unclear. Here I report the use of mass spectrometry in the identification of intermediate species in the biosynthesis and D-alanylation of lipoteichoic acid: the glycolipid anchor, nascent lipoteichoic acid primer with one phosphoglycerol unit, as well as mono- and di-alanylated forms of the lipoteichoic acid primer. Monitoring these species as well as the recently reported D-alanyl-phosphatidyl glycerol would aid in shedding light on the mechanism of the D-alanylation pathway of lipoteichoic acid.

Post-translational control of vegetative cell separation enzymes through a direct interaction with specific inhibitor IseA in Bacillus subtilis

2008 ◽  
Vol 70 (1) ◽  
pp. 168-182 ◽  
Author(s):  
Hiroki Yamamoto ◽  
Masayuki Hashimoto ◽  
Yuhei Higashitsuji ◽  
Hiroyuki Harada ◽  
Nozomi Hariyama ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Translational Control ◽  
Cell Separation ◽  
Vegetative Cell ◽  
Specific Inhibitor ◽  
Direct Interaction

scholarly journals Expression of the σF-Directed csfB Locus Prevents Premature Appearance of σG Activity during Sporulation of Bacillus subtilis

2007 ◽  
Vol 189 (23) ◽  
pp. 8754-8757 ◽  
Author(s):  
Vasant K. Chary ◽  
Panagiotis Xenopoulos ◽  
Patrick J. Piggot
Keyword(s):  
Bacillus Subtilis ◽  
Spore Formation ◽  
Artificial Induction

ABSTRACT During sporulation, σG becomes active in the prespore upon the completion of engulfment. We show that the inactivation of the σF-directed csfB locus resulted in premature activation of σG. CsfB exerted control distinct from but overlapping with that exerted by LonA to prevent inappropriate σG activation. The artificial induction of csfB severely compromised spore formation.

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