The Essential Cell Division Protein FtsN Interacts with the Murein (Peptidoglycan) Synthase PBP1B in Escherichia coli

Müller, Patrick; Ewers, Christa; Bertsche, Ute; Anstett, Maria; Kallis, Tanja; Breukink, Eefjan; Fraipont, Claudine; Terrak, Mohammed; Nguyen-Distèche, Martine; Vollmer, Waldemar

doi:10.1074/jbc.m706390200

Cited by 139 publications

(169 citation statements)

References 46 publications

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“…This hypothesis is consistent with recent work demonstrating that (i) PBP1B interacts with PBP3 (Bertsche et al, 2006) (ii) PBP1B is preferentially recruited to the mature divisome with its regulator LpoB (Fig. 1); OM-anchored LpoB localization in the divisome depends on the presence of PBP3 during septal PG synthesis and the complex PBP1B/LpoB could thus contribute to OM constriction after PBP3 intervention (Paradis-Bleau et al, 2010;Typas et al, 2010) (iii) PBP1B interacts with FtsN (Muller et al, 2007), which is positioned at the division site after PBP3 but just prior to cell constriction (Rico et al, 2010), and is essential for septation and Tol-Pal complex localization to the division site (Gerding et al, 2007). We can envisage that PBP3 intervention finishes at the moment of FtsN intervention.…”

Section: Resultssupporting

confidence: 79%

New Insights into the Final Stage of Bacterial Cell Division

Péhau‐Arnaudet¹,

Joseleau‐Petit²,

Arluison³

2012

AJMCB

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In Escherichia coli, cytokinesis is driven through the formation of the divisome, envelope invagination, peptidoglycan synthesis/degradation and daughter-cell separation. Although many proteins are implicated, the order and time needed for each involved protein's intervention remains ambiguous. One protein, Penicillin-Binding-Protein PBP3 (FtsI), seems particularly important for this process. In this paper, we describe a reproducible protocol to observe E. coli K-12 by Transmission Electron Cryo-Microscopy (cryo-TEM) on whole cells. We observed IM and OM invaginations during cell division and then analyzed the effects of PBP3 inhibitors on membrane-constrictive processes. Our results suggest that synthesis and invagination of the inner membrane can be completely dissociated from those of the outer membrane during a short time at a specific moment of the bacterial cell cycle, leading to the formation of two separate cytoplasms. These results help clarify the timing of PBP3 action during the bacterial cell cycle.

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Section: Resultssupporting

confidence: 79%

New Insights into the Final Stage of Bacterial Cell Division

Péhau‐Arnaudet¹,

Joseleau‐Petit²,

Arluison³

2012

AJMCB

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“…In E. coli, PBP1b interacts with different proteins during the course of cellular growth and division. For example, MltA, the membranebound lytic transglycosylase, interacts with PBP1b and participates in the peptidoglycan processing during cell elongation and cell division (17); PBP3, a transpeptidase catalyzing the formation of cross-linked peptidoglycan, interacts with PBP1b for peptidoglycan synthesis during cell division (18); FtsN, the essential cell division protein that can interact with PBP1b, may play a role in stabilizing the divisome during cell division (19). We hence tested the idea that whether UB2H serves as the binding domain in PBP1b for the interaction with different binding partners.…”

Section: Resultsmentioning

confidence: 99%

Crystal structure of the membrane-bound bifunctional transglycosylase PBP1b from Escherichia coli

Sung

Lai²,

Huang³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

161

180

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Drug-resistant bacteria have caused serious medical problems in recent years, and the need for new antibacterial agents is undisputed. Transglycosylase, a multidomain membrane protein essential for cell wall synthesis, is an excellent target for the development of new antibiotics. Here, we determined the X-ray crystal structure of the bifunctional transglycosylase penicillin-binding protein 1b (PBP1b) from Escherichia coli in complex with its inhibitor moenomycin to 2.16-Å resolution. In addition to the transglycosylase and transpeptidase domains, our structure provides a complete visualization of this important antibacterial target, and reveals a domain for protein-protein interaction and a transmembrane helix domain essential for substrate binding, enzymatic activity, and membrane orientation.antibacterial development ͉ antibiotic resistance ͉ membrane protein structure ͉ peptidoglycan synthesis ͉ protein-protein interaction I n the last decade, the prevalence and occasional outbreaks of drug-resistant bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE), have posed appalling hurdles in the treatment of bacterial infections (1, 2). New antibacterial agents are, as a result, in desperate demand to combat these pernicious antibiotic-resistant problems that can otherwise cause life-or-death struggles.Bacteria cell wall is a mesh-like structure of cross-linked peptidoglycan, which is essential to scaffold the cytoplasmic membrane and to maintain structural integrity of the cell (3). Cell wall synthesis at the membrane surface is mainly carried out by the membrane-bound enzymes, transpeptidases and transglycosylases, and inhibitors of the transpeptidase are among the most popular antibiotics in clinical use today (3).Escherichia coli PBP1b is a bifunctional transglycosylase, also known as peptidoglycan glycosyltransferase or murein synthase. It contains a transmembrane (TM) helix, 2 enzymatic domains [transglycosylase (TG) and transpeptidase (TP)] (4), and a domain composed of Ϸ100 aa residues between TM and TG with unknown structure and functionality (Fig. 1B). For Ͼ50 years, TP has been the main target for 2 most important classes of antibiotics: ␤-lactams (e.g., penicillin and methicillin) and glycopeptides (e.g., vancomycin). Not too long after they were introduced, resistant bacteria had emerged rapidly and caused serious medical problems. In contrast, resistant strains against moenomycin, the only natural inhibitor to TG from Streptomyces, have rarely been found. The development of new antibiotics against TG has been highly anticipated (5), and not until recently have the molecular structures of TG been available, even with the TM structure undefined.Two crystal structures of transglycosylase, a bifunctional transglycosylase from S. aureus (referred to as SaPBP2) and a transglycosylase domain from Aquifex aeolicus (referred to as AaPGT), have been determined recently with their TM domain or TM and TP domains removed, respectively (6-8). These structur...

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“…Understanding how these proteins are targeted to that site is important for understanding how division is spatially and temporally regulated. The division apparatus is generally thought of as a multiprotein complex (or a collection of smaller complexes) whose assembly is driven, for the most part, by protein-protein interactions (23,(31)(32)(33)(34)(35)(36). However proteins that contain a small PG-binding domain known as a "SPOR domain" are thought to be recruited by binding to septal PG (15)(16)(17).…”

Section: Discussionmentioning

confidence: 99%

Bacterial SPOR domains are recruited to septal peptidoglycan by binding to glycan strands that lack stem peptides

Yahashiri

Jorgenson

Weiss

2015

Proc. Natl. Acad. Sci. U.S.A.

102

142

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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.A defining feature of bacteria is the peptidoglycan (PG) cell wall or "sacculus" that confers cell shape, protects the cell against lysis caused by turgor pressure, and is the ultimate target of many clinically relevant antibiotics, including β-lactams and vancomycin (1-4). The structure of PG is characterized by a network of glycan strands connected at regular intervals by oligopeptide cross-links (Fig. 1). The glycans are built from a repeating disaccharide of β-1,4-linked N-acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc), from which branch the oligopeptides that mediate cross-linking. During cell division, a new PG wall is assembled between the incipient daughter cells (5, 6), and subsequent cell separation depends on the activity of a collection of PG hydrolases that cleave various bonds in the PG mesh (7).In the model Gram-negative bacterium Escherichia coli, daughter-cell separation is driven primarily by three cell-wall amidases (8-10). These enzymes cleave the amide bond that joins the lactyl group of the MurNAc to the α-amino group of the first amino acid (L-Ala) of the stem peptide, releasing as products free oligopeptides and "denuded" glycan strands (Fig. 1). Lytic transglycosylases (LTs) that cleave the MurNAc-GlcNAc glycosidic bond and endopeptidases that cleave stem peptides make minor but still significant contributions to daughter-cell separation (9, 11). In Bacillus subtilis, a model Gram-positive species, the enzymology of daughter-cell separation is not as well characterized. The...

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The Essential Cell Division Protein FtsN Interacts with the Murein (Peptidoglycan) Synthase PBP1B in Escherichia coli

Cited by 139 publications

References 46 publications

New Insights into the Final Stage of Bacterial Cell Division

New Insights into the Final Stage of Bacterial Cell Division

Crystal structure of the membrane-bound bifunctional transglycosylase PBP1b from Escherichia coli

Bacterial SPOR domains are recruited to septal peptidoglycan by binding to glycan strands that lack stem peptides

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