LocZ Is a New Cell Division Protein Involved in Proper Septum Placement in Streptococcus pneumoniae

Holečková, Nela; Doubravová, Linda; Massidda, Orietta; Molle, Virginie; Buriánková, Karolína; Benada, Oldřich; Kofroňová, Olga; Ulrych, Aleš; Branný, Pavel

doi:10.1128/mbio.01700-14

Cited by 87 publications

(140 citation statements)

References 63 publications

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“…Markerless deletions of the division genes, mapZ (alternative name, locZ) and divIVA, fused short (20-amino acid) stretches from the beginning and end of the genes to maintain possible translational coupling (see Tables S1 and S2). The phenotypes of the markerless ⌬mapZ and ⌬divIVA mutants were similar to those reported previously for other mapZ and divIVA mutants (41)(42)(43).…”

Section: Methodssupporting

confidence: 86%

“…9, left). This result was somewhat unexpected, given that MapZ has been postulated to be required for FtsZ positioning (42,85,86). Consistent with their relatively normal cell morphologies, long pulse-chase-new labeling of these cells revealed only occasional misplaced PG synthesis, which is a surrogate indicator of FtsZ location (33), in the ⌬mapZ mutant compared to that in the parent strain (Fig.…”

Section: Figmentioning

confidence: 83%

“…9). MapZ (also called LocZ) is a recently reported protein that has been hypothesized to direct FtsZ to the equators of future dividing daughter cells (42,85,86). Using FtsZ-green fluorescent protein (GFP) fusions, FtsZ was reported to be greatly mislocalized in ⌬mapZ mutants, which showed severe morphology defects (42,86).…”

Section: Figmentioning

confidence: 99%

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Minimal Peptidoglycan (PG) Turnover in Wild-Type and PG Hydrolase and Cell Division Mutants of Streptococcus pneumoniae D39 Growing Planktonically and in Host-Relevant Biofilms

et al. 2015

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We determined whether there is turnover of the peptidoglycan (PG) cell wall of the ovococcus bacterial pathogen Streptococcus pneumoniae (pneumococcus). Pulse-chase experiments on serotype 2 strain D39 radiolabeled with N-acetylglucosamine revealed little turnover and release of PG breakdown products during growth compared to published reports of PG turnover in Bacillus subtilis. PG dynamics were visualized directly by long-pulse-chase-new-labeling experiments using two colors of fluorescent D-amino acid (FDAA) probes to microscopically detect regions of new PG synthesis. Consistent with minimal PG turnover, hemispherical regions of stable "old" PG persisted in D39 and TIGR4 (serotype 4) cells grown in rich brain heart infusion broth, in D39 cells grown in chemically defined medium containing glucose or galactose as the carbon source, and in D39 cells grown as biofilms on a layer of fixed human epithelial cells. In contrast, B. subtilis exhibited rapid sidewall PG turnover in similar FDAA-labeling experiments. High-performance liquid chromatography (HPLC) analysis of biochemically released peptides from S. pneumoniae PG validated that FDAAs incorporated at low levels into pentamer PG peptides and did not change the overall composition of PG peptides. PG dynamics were also visualized in mutants lacking PG hydrolases that mediate PG remodeling, cell separation, or autolysis and in cells lacking the MapZ and DivIVA division regulators. In all cases, hemispheres of stable old PG were maintained. In PG hydrolase mutants exhibiting aberrant division plane placement, FDAA labeling revealed patches of inert PG at turns and bulge points. We conclude that growing S. pneumoniae cells exhibit minimal PG turnover compared to the PG turnover in rod-shaped cells. IMPORTANCEPG cell walls are unique to eubacteria, and many bacterial species turn over and recycle their PG during growth, stress, colonization, and virulence. Consequently, PG breakdown products serve as signals for bacteria to induce antibiotic resistance and as activators of innate immune responses. S. pneumoniae is a commensal bacterium that colonizes the human nasopharynx and opportunistically causes serious respiratory and invasive diseases. The results presented here demonstrate a distinct demarcation between regions of old PG and regions of new PG synthesis and minimal turnover of PG in S. pneumoniae cells growing in culture or in host-relevant biofilms. These findings suggest that S. pneumoniae minimizes the release of PG breakdown products by turnover, which may contribute to evasion of the innate immune system. P eptidoglycan (PG) biosynthesis and placement are dynamic processes that determine the shapes, sizes, chaining, and resistance to turgor of bacterial cells (1-6). In Gram-positive bacteria, PG also serves as the scaffolding for covalent attachment of surface wall teichoic acid, capsule, and sortase-attached proteins (7-9). The seminal work of Park and Uehara demonstrated that PG is rapidly turned over and the breakdown components recycled in...

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Section: Methodssupporting

confidence: 86%

Section: Figmentioning

confidence: 83%

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Minimal Peptidoglycan (PG) Turnover in Wild-Type and PG Hydrolase and Cell Division Mutants of Streptococcus pneumoniae D39 Growing Planktonically and in Host-Relevant Biofilms

et al. 2015

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“…Ovococcal S. pneumoniae lacks a nucleoid occlusion system and has no Min-system (20,21). Recently, MapZ (or LocZ) was proposed to be a division site selector in S. pneumoniae (22,23). This protein localizes early at new cell division sites and positions FtsZ by a direct protein-protein interaction (22).…”

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confidence: 99%

Chromosome segregation drives division site selection in Streptococcus pneumoniae

Raaphorst

Kjos

Veening

2017

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

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Accurate spatial and temporal positioning of the tubulin-like protein FtsZ is key for proper bacterial cell division. Streptococcus pneumoniae (pneumococcus) is an oval-shaped, symmetrically dividing opportunistic human pathogen lacking the canonical systems for division site control (nucleoid occlusion and the Min-system). Recently, the early division protein MapZ was identified and implicated in pneumococcal division site selection. We show that MapZ is important for proper division plane selection; thus, the question remains as to what drives pneumococcal division site selection. By mapping the cell cycle in detail, we show that directly after replication both chromosomal origin regions localize to the future cell division sites, before FtsZ. Interestingly, Z-ring formation occurs coincidently with initiation of DNA replication. Perturbing the longitudinal chromosomal organization by mutating the condensin SMC, by CRISPR/Cas9-mediated chromosome cutting, or by poisoning DNA decatenation resulted in mistiming of MapZ and FtsZ positioning and subsequent cell elongation. Together, we demonstrate an intimate relationship between DNA replication, chromosome segregation, and division site selection in the pneumococcus, providing a simple way to ensure equally sized daughter cells.

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“…DivIVA, PBP1a, and PBP2x (39, 41) but after LocZ (46). Given the localization profile and that S. pneumoniae FtsA also interacts directly with FtsZ, it was hypothesized that the two proteins arrive at midcell at the same time and colocalize during the cell cycle (35).…”

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confidence: 99%

Roles of the Essential Protein FtsA in Cell Growth and Division in Streptococcus pneumoniae

et al. 2017

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Streptococcus pneumoniae is an ovoid-shaped Gram-positive bacterium that grows by carrying out peripheral and septal peptidoglycan (PG) synthesis, analogous to model bacilli, such as Escherichia coli and Bacillus subtilis. In the model bacilli, FtsZ and FtsA proteins assemble into a ring at midcell and are dedicated to septal PG synthesis but not peripheral PG synthesis; hence, inactivation of FtsZ or FtsA results in long filamentous cells unable to divide. Here, we demonstrate that FtsA and FtsZ colocalize at midcell in S. pneumoniae and that partial depletion of FtsA perturbs septum synthesis, resulting in elongated cells with multiple FtsZ rings that fail to complete septation. Unexpectedly, complete depletion of FtsA resulted in the delocalization of FtsZ rings and ultimately cell ballooning and lysis. In contrast, depletion or deletion of gpsB and sepF, which in B. subtilis are synthetically lethal with ftsA, resulted in enlarged and elongated cells with multiple FtsZ rings, with deletion of sepF mimicking partial depletion of FtsA. Notably, cell ballooning was not observed, consistent with later recruitment of these proteins to midcell after Z-ring assembly. The overproduction of FtsA stimulates septation and suppresses the cell division defects caused by the deletion of sepF and gpsB under some conditions, supporting the notion that FtsA shares overlapping functions with GpsB and SepF at later steps in the division process. Our results indicate that, in S. pneumoniae, both GpsB and SepF are involved in septal PG synthesis, whereas FtsA and FtsZ coordinate both peripheral and septal PG synthesis and are codependent for localization at midcell.IMPORTANCE Streptococcus pneumoniae (pneumococcus) is a clinically important human pathogen for which more therapies against unexploited essential targets, like cell growth and division proteins, are needed. Pneumococcus is an ovoid-shaped Gram-positive bacterium with cell growth and division properties that have important distinctions from those of rod-shaped bacteria. Gaining insights into these processes can thus provide valuable information to develop novel antimicrobials. Whereas rods use distinctly localized protein machines at different cellular locations to synthesize peripheral and septal peptidoglycans, we present evidence that S. pneumoniae organizes these two machines at a single location in the middle of dividing cells. Here, we focus on the properties of the actin-like protein FtsA as an essential orchestrator of peripheral and septal growth in this bacterium.KEYWORDS FtsA, Gram-positive cocci, Streptococcus pneumoniae, cell division

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LocZ Is a New Cell Division Protein Involved in Proper Septum Placement in Streptococcus pneumoniae

Cited by 87 publications

References 63 publications

Minimal Peptidoglycan (PG) Turnover in Wild-Type and PG Hydrolase and Cell Division Mutants of Streptococcus pneumoniae D39 Growing Planktonically and in Host-Relevant Biofilms

Minimal Peptidoglycan (PG) Turnover in Wild-Type and PG Hydrolase and Cell Division Mutants of Streptococcus pneumoniae D39 Growing Planktonically and in Host-Relevant Biofilms

Chromosome segregation drives division site selection in Streptococcus pneumoniae

Roles of the Essential Protein FtsA in Cell Growth and Division in Streptococcus pneumoniae

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