title: Post-translational control of vegetative cell separation enzymes through a direct interaction with specific inhibitor IseA in Bacillus subtilis.
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.
authors.affiliation: Experimental Farm, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda-shi, Nagano 386-8567, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Masayuki
authors.initials: M
authors.lastname: Hashimoto
authors.forename: Yuhei
authors.initials: Y
authors.lastname: Higashitsuji
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Harada
authors.forename: Nozomi
authors.initials: N
authors.lastname: Hariyama
authors.forename: Lisa
authors.initials: L
authors.lastname: Takahashi
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Iwashita
authors.forename: Seika
authors.initials: S
authors.lastname: Ooiwa
authors.forename: Junichi
authors.initials: J
authors.lastname: Sekiguchi