PMID:21472954

Citation	Olrichs, NK, Aarsman, ME, Verheul, J, Arnusch, CJ, Martin, NI, Hervé, M, Vollmer, W, de Kruijff, B, Breukink, E and den Blaauwen, T (2011) A novel in vivo cell-wall labeling approach sheds new light on peptidoglycan synthesis in Escherichia coli.Chembiochem 12:1124-33
Abstract	Peptidoglycan synthesis and turnover in relation to cell growth and division has been studied by using a new labeling method. This method involves the incorporation of fluorescently labeled peptidoglycan precursors into the cell wall by means of the cell-wall recycling pathway. We show that Escherichia coli is able to import exogenous added murein tripeptide labeled with N-7-nitro-2,1,3-benzoxadiazol-4-yl (AeK-NBD) into the cytoplasm where it enters the peptidoglycan biosynthesis route, resulting in fluorescent labels specifically located in the cell wall. When wild-type cells were grown in the presence of the fluorescent peptide, peptidoglycan was uniformly labeled in cells undergoing elongation. Cells in the process of division displayed a lack of labeled peptidoglycan at mid-cell. Analysis of labeling patterns in cell division mutants showed that the occurrence of unlabeled peptidoglycan is dependent on the presence of FtsZ, but independent of FtsQ and FtsI. Accumulation of fluorescence at the division sites of a triple amidase mutant (ΔamiABC) revealed that AeK-NBD is incorporated into septal peptidoglycan. AmiC was shown to be involved in the rapid removal of labeled peptidoglycan side chains at division sites in wild-type cells. Because septal localization of AmiC is dependent on FtsQ and FtsI, this points to the presence of another peptidoglycan hydrolase activity directly dependent on FtsZ.
Links	PubMed Online version:10.1002/cbic.201000552
Keywords	Cell Wall; Escherichia coli; Peptidoglycan; Staining and Labeling
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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: W3899 NCBI Taxon ID: 83333	Genotype of Reference Strain: amiABC Genotype of Experimental Strain : amiABC(del)	Reference Condition:					when Supplying cells with the cell-wall precursor murein tripeptide (AeK) labeled with NBD accumulation of AeK-NBD occurs at division site indicating labeled precursors are utilized for septal peptidoglycan synthesis and amiC activity is mainly responsible for the rapid removal of labeled murein peptides at division sites as indicated by double mutant amiAB(del), figure 7.
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