Controlling septum thickness by a large protein ring

AbstractGram-positive bacteria divide by forming a thick cross wall. How the thickness of this septal wall is controlled is unknown. In this type of bacteria, the key cell division protein FtsZ is anchored to the cell membrane by two proteins, FtsA and SepF. We have isolated SepF homologues from different bacterial species and found that they all polymerize into large protein rings with diameters varying from 19 to 41 nm. Importantly, these values correlated well with the thickness of their septa. To test whether ring diameter determines septal thickness, we tried to construct different SepF chimeras with the purpose to manipulate the diameter of the SepF protein ring. This was indeed possible and confirmed that the conserved core domain of SepF determines ring diameter. Importantly, when SepF chimeras with a smaller diameter were expressed in the bacterial host Bacillus subtilis, the thickness of its septa also became smaller. These results strongly support a model in which septal thickness is controlled by curved molecular clamps formed by SepF polymers attached to the leading edge of nascent septa. This also implies that the intrinsic shape of a protein polymer can function as a mould to shape the cell wall.Significance StatementMany bacteria form a thick cell wall and divide by forming a cross wall. How they control the thickness of their cell wall and cross wall is unknown. In this study we show that in these bacteria the cell division protein SepF forms very large protein rings with diameters that correspond to the diameter of their cross walls. Importantly, when we reduced the diameter of SepF rings in the bacterial host Bacillus subtilis the cross wall also became thinner. These results provide strong evidence that a large protein ring can function as a mould to control the thickness of the cell wall that divides these bacterial cells.

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Control of septum thickness by the curvature of SepF polymers

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2002635118 ◽

2020 ◽

Vol 118 (2) ◽

pp. e2002635118

Author(s):

Michaela Wenzel ◽

Ilkay N. Celik Gulsoy ◽

Yongqiang Gao ◽

Zihao Teng ◽

Joost Willemse ◽

...

Keyword(s):

Bacterial Species ◽

Leading Edge ◽

Cross Wall ◽

Septal Wall ◽

Gram Positive Bacteria ◽

Septal Thickness ◽

Ring Diameter ◽

Intrinsic Shape ◽

Different Diameters ◽

Conserved Core

Gram-positive bacteria divide by forming a thick cross wall. How the thickness of this septal wall is controlled is unknown. In this type of bacteria, the key cell division protein FtsZ is anchored to the cell membrane by two proteins, FtsA and/or SepF. We have isolated SepF homologs from different bacterial species and found that they all polymerize into large protein rings with diameters varying from 19 to 44 nm. Interestingly, these values correlated well with the thickness of their septa. To test whether ring diameter determines septal thickness, we tried to construct different SepF chimeras with the purpose to manipulate the diameter of the SepF protein ring. This was indeed possible and confirmed that the conserved core domain of SepF regulates ring diameter. Importantly, when SepF chimeras with different diameters were expressed in the bacterial hostBacillus subtilis, the thickness of its septa changed accordingly. These results strongly support a model in which septal thickness is controlled by curved molecular clamps formed by SepF polymers attached to the leading edge of nascent septa. This also implies that the intrinsic shape of a protein polymer can function as a mold to shape the cell wall.

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Cell division protein DivIB influences the Spo0J/Soj system of chromosome segregation in Bacillus subtilis

Molecular Microbiology ◽

10.1111/j.1365-2958.2004.04399.x ◽

2004 ◽

Vol 55 (2) ◽

pp. 349-367 ◽

Cited By ~ 22

Author(s):

Gonçalo Real ◽

Sabine Autret ◽

Elizabeth J. Harry ◽

Jeffery Errington ◽

Adriano O. Henriques

Keyword(s):

Bacillus Subtilis ◽

Cell Division ◽

Chromosome Segregation ◽

Cell Division Protein

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Bacteriophage SP01 Gene Product 56 Inhibits Bacillus subtilis Cell Division by Interacting with FtsL and Disrupting Pbp2B and FtsW Recruitment

Journal of Bacteriology ◽

10.1128/jb.00463-20 ◽

2020 ◽

Vol 203 (2) ◽

pp. e00463-20

Author(s):

Amit Bhambhani ◽

Isabella Iadicicco ◽

Jules Lee ◽

Syed Ahmed ◽

Max Belfatto ◽

...

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Cell Division ◽

Gene Product ◽

Fluorescent Protein ◽

Lytic Bacteriophage ◽

Cell Wall Synthesis ◽

Content Type ◽

Ftsz Ring ◽

Green Fluorescent

ABSTRACTPrevious work identified gene product 56 (gp56), encoded by the lytic bacteriophage SP01, as being responsible for inhibition of Bacillus subtilis cell division during its infection. Assembly of the essential tubulin-like protein FtsZ into a ring-shaped structure at the nascent site of cytokinesis determines the timing and position of division in most bacteria. This FtsZ ring serves as a scaffold for recruitment of other proteins into a mature division-competent structure permitting membrane constriction and septal cell wall synthesis. Here, we show that expression of the predicted 9.3-kDa gp56 of SP01 inhibits later stages of B. subtilis cell division without altering FtsZ ring assembly. Green fluorescent protein-tagged gp56 localizes to the membrane at the site of division. While its localization does not interfere with recruitment of early division proteins, gp56 interferes with the recruitment of late division proteins, including Pbp2b and FtsW. Imaging of cells with specific division components deleted or depleted and two-hybrid analyses suggest that gp56 localization and activity depend on its interaction with FtsL. Together, these data support a model in which gp56 interacts with a central part of the division machinery to disrupt late recruitment of the division proteins involved in septal cell wall synthesis.IMPORTANCE Studies over the past decades have identified bacteriophage-encoded factors that interfere with host cell shape or cytokinesis during viral infection. The phage factors causing cell filamentation that have been investigated to date all act by targeting FtsZ, the conserved prokaryotic tubulin homolog that composes the cytokinetic ring in most bacteria and some groups of archaea. However, the mechanisms of several phage factors that inhibit cytokinesis, including gp56 of bacteriophage SP01 of Bacillus subtilis, remain unexplored. Here, we show that, unlike other published examples of phage inhibition of cytokinesis, gp56 blocks B. subtilis cell division without targeting FtsZ. Rather, it utilizes the assembled FtsZ cytokinetic ring to localize to the division machinery and to block recruitment of proteins needed for septal cell wall synthesis.

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Ultrastructure of a marine psychrophilic Vibrio

Canadian Journal of Microbiology ◽

10.1139/m70-174 ◽

1970 ◽

Vol 16 (11) ◽

pp. 1027-1031 ◽

Cited By ~ 7

Author(s):

S. F. Kennedy ◽

R. R. Colwell ◽

G. B. Chapman

Keyword(s):

Electron Microscopy ◽

Cell Wall ◽

Plasma Membrane ◽

Cell Division ◽

Wall Material ◽

Cross Wall ◽

Growth Stages ◽

Culture Age ◽

Primary Mechanism ◽

Age Studies

The structure of Vibrio marinus strain PS-207 was studied by both phase and electron microscopy. It was found to possess a trilaminar plasma membrane and cell wall. Membrane-bounded subunits containing DNA-like material were found dispersed throughout the cytoplasm. Giant round forms or "macrospheres" were observed in all growth stages. The size, shape, and construction of the "macrospheres" showed some variation, but could not be related to culture age. Studies of cell division in V. marinus strain PS-207 indicate the primary mechanism to be a synthesis and centripetal deposition of plasma membrane with a concomitant or subsequent synthesis and centripetal deposition of cross wall material.

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FtsW exhibits distinct processive motions driven by either septal cell wall synthesis or FtsZ treadmilling in E. coli

10.1101/850073 ◽

2019 ◽

Cited By ~ 8

Author(s):

Xinxing Yang ◽

Ryan McQuillen ◽

Zhixin Lyu ◽

Polly Phillips-Mason ◽

Ana De La Cruz ◽

...

Keyword(s):

Cell Wall ◽

Cell Division ◽

Single Molecule ◽

Bacterial Cell ◽

Spatial Information ◽

Bacterial Species ◽

E Coli ◽

Slow Moving

AbstractDuring bacterial cell division, synthesis of new septal peptidoglycan (sPG) is crucial for successful cytokinesis and cell pole morphogenesis. FtsW, a SEDS (Shape, Elongation, Division and Sporulation) family protein and an indispensable component of the cell division machinery in all walled bacterial species, was recently identified in vitro as a new monofunctional peptidoglycan glycosyltransferase (PGTase). FtsW and its cognate monofunctional transpeptidase (TPase) class B penicillin binding protein (PBP3 or FtsI in E. coli) may constitute the essential, bifunctional sPG synthase specific for new sPG synthesis. Despite its importance, the septal PGTase activity of FtsW has not been documented in vivo. How its activity is spatiotemporally regulated in vivo has also remained unknown. Here we investigated the septal PGTase activity and dynamics of FtsW in E. coli cells using a combination of single-molecule imaging and genetic manipulations. We show that FtsW exhibits robust activity to incorporate an N-acetylmuramic acid analog at septa in the absence of other known PGTases, confirming FtsW as the essential septum-specific PGTase in vivo. Notably, we identified two populations of processive moving FtsW molecules at septa. A fast-moving population is driven by the treadmilling dynamics of FtsZ and independent of sPG synthesis. A slow-moving population is driven by active sPG synthesis and independent of FtsZ’s treadmilling dynamics. We further identified that FtsN, a potential sPG synthesis activator, plays an important role in promoting the slow-moving, sPG synthesis-dependent population. Our results support a two-track model, in which inactive sPG synthase molecules follow the fast treadmilling “Z-track” to be distributed along the septum; FtsN promotes their release from the “Z-track” to become active in sPG synthesis on the slow “sPG-track”. This model explains how the spatial information is integrated into the regulation of sPG synthesis activity and suggests a new mechanistic framework for the spatiotemporal coordination of bacterial cell wall constriction.

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Transcription factor Spo0A switches the localization of the cell division protein FtsZ from a medial to a bipolar pattern in Bacillus subtilis.

Genes & Development ◽

10.1101/gad.10.4.478 ◽

1996 ◽

Vol 10 (4) ◽

pp. 478-488 ◽

Cited By ~ 193

Author(s):

P A Levin ◽

R Losick

Keyword(s):

Transcription Factor ◽

Bacillus Subtilis ◽

Cell Division ◽

Cell Division Protein

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DivIVA Is Required for Polar Growth in the MreB-Lacking Rod-Shaped Actinomycete Corynebacterium glutamicum

Journal of Bacteriology ◽

10.1128/jb.01934-07 ◽

2008 ◽

Vol 190 (9) ◽

pp. 3283-3292 ◽

Cited By ~ 94

Author(s):

Michal Letek ◽

Efrén Ordóñez ◽

José Vaquera ◽

William Margolin ◽

Klas Flärdh ◽

...

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Cell Division ◽

Streptococcus Pneumoniae ◽

Corynebacterium Glutamicum ◽

Chromosome Segregation ◽

Polar Growth ◽

Cell Wall Synthesis ◽

Peptidoglycan Synthesis ◽

Polarized Cell Growth

ABSTRACT The actinomycete Corynebacterium glutamicum grows as rod-shaped cells by zonal peptidoglycan synthesis at the cell poles. In this bacterium, experimental depletion of the polar DivIVA protein (DivIVACg) resulted in the inhibition of polar growth; consequently, these cells exhibited a coccoid morphology. This result demonstrated that DivIVA is required for cell elongation and the acquisition of a rod shape. DivIVA from Streptomyces or Mycobacterium localized to the cell poles of DivIVACg-depleted C. glutamicum and restored polar peptidoglycan synthesis, in contrast to DivIVA proteins from Bacillus subtilis or Streptococcus pneumoniae, which localized at the septum of C. glutamicum. This confirmed that DivIVAs from actinomycetes are involved in polarized cell growth. DivIVACg localized at the septum after cell wall synthesis had started and the nucleoids had already segregated, suggesting that in C. glutamicum DivIVA is not involved in cell division or chromosome segregation.

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