The monofunctional glycosyltransferase of Escherichia coli localizes to the cell division site and interacts with penicillin-binding protein 3, FtsW, and FtsN - PubMed (original) (raw)

The monofunctional glycosyltransferase of Escherichia coli localizes to the cell division site and interacts with penicillin-binding protein 3, FtsW, and FtsN

Adeline Derouaux et al. J Bacteriol. 2008 Mar.

Abstract

The monofunctional peptidoglycan glycosyltransferase (MtgA) catalyzes glycan chain elongation of the bacterial cell wall. Here we show that MtgA localizes at the division site of Escherichia coli cells that are deficient in PBP1b and produce a thermosensitive PBP1a and is able to interact with three constituents of the divisome, PBP3, FtsW, and FtsN, suggesting that MtgA may play a role in peptidoglycan assembly during the cell cycle in collaboration with other proteins.

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Figures

FIG. 1.

FIG. 1.

Localization of GFP-MtgA in wild-type ponA+ ponB+ strain LMC500 (A) and ponA(ts) ponB strain EJ801 (B) grown in LB medium at 28°C, both harboring pDML2005. The arrows point to cells with fluorescent signals at the division site. Photographs were taken with a cooled AxioCam MRm (Zeiss) mounted on an Zeiss Axio Imager.Z1 fluorescent microscope through a EC Plan-Neofluar 100 × 1.3 oil immersion objective in bright field and fluorescence using filter set 37 (400- to 500-nm band pass excitation and 460- to 560-nm band pass emission; Zeiss). Images were analyzed using the AxioVision Rel. 4.5 (Zeiss) software as previously described (24). Bar, 5 μm.

FIG. 2.

FIG. 2.

Interaction between MtgA and PBP3, FtsW, FtsN, or itself by the Cya bacterial two-hybrid assay. DHM1 transformants producing T18-T25 (1), T18-MtgA-T25 (2), T18-T25-MtgA (3), T18-MtgA-T25-Lpp-PBP3 (4), T18-Lpp-PBP3-T25-MtgA (5), T18-PBP1B-T25-PBP3 (6), T18-PBP3-T25-PBP1B (7), T18-MtgA-T25-PBP3 (8), T18-PBP3-T25-MtgA (9), T18-MtgA-T25-FtsN (10), T18-FstN-T25-MtgA (11), T18-MtgA-T25-FstW (12), T18-FtsW-T25-MtgA (13), and T18-MtgA-T25-MtgA (14) were used. (A) Transformants were grown at 30°C for 30 h on LB agar plates containing 0.5 mM IPTG, 40 μg/ml X-Gal, 50 μg/ml ampicillin, and 20 μg/ml chloramphenicol. (B) Quantitative analysis of the β-galactosidase activity from transformants grown in LB medium in the presence of 0.4 mM IPTG for 16 h at 30°C was carried out as described previously (17). Data are averages of three independent experiments.

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