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

Hiroki Yamamoto, Masayuki Hashimoto, Yuhei Higashitsuji, Hiroyuki Harada, Nozomi Hariyama, Lisa Takahashi, Tomoaki Iwashita, Seika Ooiwa, Junichi Sekiguchi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Three D,L-endopeptidases, LytE, LytF and CwlS, are involved in the vegetative cell separation in Bacillus subtilis. A novel cell surface protein, IseA, inhibits the cell wall lytic activities of these D,L-endopeptidases in vitro, and IseA negatively regulates the cell separation enzymes at the post-translational level. Immunofluorescence microscopy indicated that the IseA-3xFLAG fusion protein was specifically localized at cell separation sites and poles on the vegetative cell surface in a similar manner of the D,L-endopeptidases. Furthermore, pull-down assay showed that IseA binds to the catalytic domain of LytF, indicating that IseA is localized on the cell surface through the catalytic domain of LytF. Overexpression of IseA caused a long-chained cell morphology in the exponential growth phase, indicating that IseA inhibits the cell separation D,L-endopeptidases in vivo. Besides, overexpression of IseA in a cwlO disruptant affected cell growth, implying that IseA is also involved in the cell elongation event. However, although IseA inhibits the activities of LytE, LytF, CwlS and CwlO in vitro, it is unlikely to inhibit CwlS and CwlO in vivo. This is the first demonstration that the cell separation event is post-translationally controlled through a direct interaction between cell separation enzymes and a specific novel inhibitor in bacteria.

Original languageEnglish
Pages (from-to)168-182
Number of pages15
JournalMolecular Microbiology
Volume70
Issue number1
DOIs
Publication statusPublished - 2008 Oct 1

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Cell Separation
Bacillus subtilis
Endopeptidases
Enzymes
Catalytic Domain
Growth
Fluorescence Microscopy
Cell Wall
Membrane Proteins
Bacteria
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Yamamoto, Hiroki ; Hashimoto, Masayuki ; Higashitsuji, Yuhei ; Harada, Hiroyuki ; Hariyama, Nozomi ; Takahashi, Lisa ; Iwashita, Tomoaki ; Ooiwa, Seika ; Sekiguchi, Junichi. / Post-translational control of vegetative cell separation enzymes through a direct interaction with specific inhibitor IseA in Bacillus subtilis. In: Molecular Microbiology. 2008 ; Vol. 70, No. 1. pp. 168-182.
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Post-translational control of vegetative cell separation enzymes through a direct interaction with specific inhibitor IseA in Bacillus subtilis. / Yamamoto, Hiroki; Hashimoto, Masayuki; Higashitsuji, Yuhei; Harada, Hiroyuki; Hariyama, Nozomi; Takahashi, Lisa; Iwashita, Tomoaki; Ooiwa, Seika; Sekiguchi, Junichi.

In: Molecular Microbiology, Vol. 70, No. 1, 01.10.2008, p. 168-182.

Research output: Contribution to journalArticle

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AU - Yamamoto, Hiroki

AU - Hashimoto, Masayuki

AU - Higashitsuji, Yuhei

AU - Harada, Hiroyuki

AU - Hariyama, Nozomi

AU - Takahashi, Lisa

AU - Iwashita, Tomoaki

AU - Ooiwa, Seika

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