Identification of SPOR Domain Amino Acids Important for Septal Localization, Peptidoglycan Binding, and a Disulfide Bond in the Cell Division Protein FtsN

Duncan, Tammi R.; Yahashiri, Atsushi; Arends, S J Ryan; Popham, David L.; Weiss, David S.

doi:10.1128/jb.00911-13

Cited by 22 publications

(36 citation statements)

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“…However, it is not yet possible to model the SPOR:PG complex to address issues such as whether the presence of stem peptides sterically blocks binding of the glycan chain to the protein. Mutagenesis of the SPOR domain from FtsN led to the unexpected discovery of a disulfide bond that is critical for the stability of the domain (51). Subsequently, Beckwith's group demonstrated that the disulfide is required for the overall stability of FtsN, so much so, in fact, that when an E. coli disulfide bond (dsb) mutant is grown anaerobically, FtsN is almost completely degraded and division fails (52).…”

Section: Spor Domains Exhibit An Rnp Fold and A Conserved Pg-binding mentioning

confidence: 99%

“…The essential nature of FtsN=s disulfide had been overlooked in previous studies of dsb mutants because molecular oxygen and/or oxidized components of the growth medium allow sufficient disulfide bond formation during aerobic growth. About 15% of SPOR domains in the sequence databases have paired cysteines, suggesting that there is a widespread connection between cell division and redox metabolism (51).…”

Section: Spor Domains Exhibit An Rnp Fold and A Conserved Pg-binding mentioning

confidence: 99%

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The SPOR Domain, a Widely Conserved Peptidoglycan Binding Domain That Targets Proteins to the Site of Cell Division

2017

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Sporulation-related repeat (SPOR) domains are small peptidoglycan (PG) binding domains found in thousands of bacterial proteins. The name "SPOR domain" stems from the fact that several early examples came from proteins involved in sporulation, but SPOR domain proteins are quite diverse and contribute to a variety of processes that involve remodeling of the PG sacculus, especially with respect to cell division. SPOR domains target proteins to the division site by binding to regions of PG devoid of stem peptides ("denuded" glycans), which in turn are enriched in septal PG by the intense, localized activity of cell wall amidases involved in daughter cell separation. This targeting mechanism sets SPOR domain proteins apart from most other septal ring proteins, which localize via protein-protein interactions. In addition to SPOR domains, bacteria contain several other PG-binding domains that can exploit features of the cell wall to target proteins to specific subcellular sites.

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Section: Spor Domains Exhibit An Rnp Fold and A Conserved Pg-binding mentioning

confidence: 99%

Section: Spor Domains Exhibit An Rnp Fold and A Conserved Pg-binding mentioning

confidence: 99%

The SPOR Domain, a Widely Conserved Peptidoglycan Binding Domain That Targets Proteins to the Site of Cell Division

2017

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“…domains from DamX and FtsN that impair septal localization in vivo also impair PG binding in vitro, indicating that PG binding is necessary for septal localization (21,22). Nevertheless, it is not known whether PG binding is sufficient for septal localization or whether denuded glycans are enriched in septal PG.…”

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confidence: 98%

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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“…FtsN was identified as a multicopy suppressor of a conditional mutation in ftsA but suppresses conditional mutations in ftsI, ftsQ, or ftsK as well (29)(30)(31). FtsN has at least four functional domains: an N-terminal cytoplasmic domain (FtsN cyto ), which contains a conserved motif responsible for interaction with FtsA; a transmembrane domain; and two periplasmic domains, the E-domain (FtsN E ), which triggers septal PG synthesis, and a C-terminal SPOR (septal peptidoglycan binding) domain, which interacts with peptidoglycan strands denuded by amidases (12,27,28,(32)(33)(34)(35)(36). FtsN E was designated as the essential domain because it can…”

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confidence: 99%

Disruption of divisome assembly rescued by FtsN–FtsA interaction in Escherichia coli

Pichoff

Lutkenhaus

2018

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

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Cell division requires the assembly of a protein complex called the divisome. The divisome assembles in a hierarchical manner, with FtsA functioning as a hub to connect the Z-ring with the rest of the divisome and FtsN arriving last to activate the machine to synthesize peptidoglycan. FtsEX arrives as the Z-ring forms and acts on FtsA to initiate recruitment of the other divisome components. In the absence of FtsEX, recruitment is blocked; however, a multitude of conditions allow FtsEX to be bypassed. Here, we find that all such FtsEX bypass conditions, as well as the bypass of FtsK, depend upon the interaction of FtsN with FtsA, which promotes the back-recruitment of the late components of the divisome. Furthermore, our results suggest that these bypass conditions enhance the weak interaction of FtsN with FtsA and its periplasmic partners so that the divisome proteins are brought to the Z-ring when the normal hierarchical pathway is disrupted.

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Identification of SPOR Domain Amino Acids Important for Septal Localization, Peptidoglycan Binding, and a Disulfide Bond in the Cell Division Protein FtsN

Cited by 22 publications

References 50 publications

The SPOR Domain, a Widely Conserved Peptidoglycan Binding Domain That Targets Proteins to the Site of Cell Division

The SPOR Domain, a Widely Conserved Peptidoglycan Binding Domain That Targets Proteins to the Site of Cell Division

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

Disruption of divisome assembly rescued by FtsN–FtsA interaction in Escherichia coli

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