title: Bacterial SPOR domains are recruited to septal peptidoglycan by binding to glycan strands that lack stem peptides.
abstract: Bacterial SPOR domains bind peptidoglycan (PG) and are thought to target proteins to the cell division site by binding to "denuded" glycan strands that lack stem peptides, but uncertainties remain, in part because septal-specific binding has yet to be studied in a purified system. Here we show that fusions of GFP to SPOR domains from the Escherichia coli cell-division proteins DamX, DedD, FtsN, and RlpA all localize to septal regions of purified PG sacculi obtained from E. coli and Bacillus subtilis. Treatment of sacculi with an amidase that removes stem peptides enhanced SPOR domain binding, whereas treatment with a lytic transglycosylase that removes denuded glycans reduced SPOR domain binding. These findings demonstrate unequivocally that SPOR domains localize by binding to septal PG, that the physiologically relevant binding site is indeed a denuded glycan, and that denuded glycans are enriched in septal PG rather than distributed uniformly around the sacculus. Accumulation of denuded glycans in the septal PG of both E. coli and B. subtilis, organisms separated by 1 billion years of evolution, suggests that sequential removal of stem peptides followed by degradation of the glycan backbone is an ancient feature of PG turnover during bacterial cell division. Linking SPOR domain localization to the abundance of a structure (denuded glycans) present only transiently during biogenesis of septal PG provides a mechanism for coordinating the function of SPOR domain proteins with the progress of cell division.
authors: Department of Microbiology, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242.
authors: Atsushi
authors: A
authors: Yahashiri
authors: Department of Microbiology, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242.
authors: Matthew A
authors: MA
authors: Jorgenson
authors: Department of Microbiology, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242 david-weiss@uiowa.edu.
authors: David S
authors: DS
authors: Weiss
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002455:Cell Division; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008954:Models, Biological; D009154:Mutation; D010455:Peptides / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism*; D011485:Protein Binding
chemical_list: C545719:DamX protein, E coli; D029968:Escherichia coli Proteins; D010455:Peptides; D010457:Peptidoglycan; D049452:Green Fluorescent Proteins
keywords: amidase; cell division; divisome; lytic transglycosylase; murein