Defining the rate-limiting processes of bacterial cytokinesis

Coltharp, Carla; Buss, Jackson; Plumer, Trevor; Xiao, Jie

doi:10.1073/pnas.1514296113

Cited by 161 publications

(277 citation statements)

References 125 publications

(192 reference statements)

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“…In B. subtilis, reduced FtsZ treadmilling rates resulted in proportionally reduced septum synthesis rates, whereas in E. coli, septum synthesis rate was not limited by treadmilling rate, although reduced treadmilling adversely affected the proper distribution and composition of septal peptidoglycan. These results are consistent with evidence that the rate of septum synthesis is limited by the activity of the PBP3 septal transpeptidase and not by the properties of the Z ring itself [16]. One possible reason for the differences between species is that the cell wall and septum of Gram-positive bacteria such as B. subtilis are thicker, demanding a higher throughput from cell wall synthesis enzymes (Fig.…”

supporting

confidence: 89%

Bacterial Division: FtsZ Treadmills to Build a Beautiful Wall

Schoenemann¹,

Margolin²

2017

Current Biology

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supporting

confidence: 89%

Bacterial Division: FtsZ Treadmills to Build a Beautiful Wall

Schoenemann¹,

Margolin²

2017

Current Biology

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“…The radial thickness was very close in magnitude to the axial width (71 nm axial width vs. 64 nm radial thickness (Holden et al 2014); Fig. 7 in (Jacq et al 2015); 99 nm axial vs. 59 nm radial (Coltharp et al 2016)). This large radial width contradicts our expectation that all pfs are tethered to the membrane by FtsA and ZipA, necesitating a Z ring that is radially one subunit thick.…”

Section: The Pf Substructure Of Z Rings – Ribbons or Scattered?mentioning

confidence: 60%

“…A recent PALM study of the Z ring in Streptococcus pneumoniae measured the axial width of the Z ring to be 95 nm early in the cell cycle, increasing to 127 nm as the Z ring constricted (Jacq et al 2015). It should be noted, however, that the most recent and comprehensive PALM studies (Coltharp et al 2016; Holden et al 2014; Jacq et al 2015) also measured the thickness of the Z ring in the radial direction. The radial thickness was very close in magnitude to the axial width (71 nm axial width vs. 64 nm radial thickness (Holden et al 2014); Fig.…”

Section: The Pf Substructure Of Z Rings – Ribbons or Scattered?mentioning

confidence: 99%

FtsZ Constriction Force – Curved Protofilaments Bending Membranes

Erickson

Osawa

2017

Subcellular Biochemistry

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FtsZ assembles in vitro into protofilaments (pfs) that are one subunit thick and ~50 subunits long. In vivo these pfs assemble further into the Z ring, which, along with accessory division proteins, constricts to divide the cell. We have reconstituted Z rings in liposomes in vitro, using pure FtsZ that was modified with a membrane targeting sequence to directly bind the membrane. This FtsZ-mts assembled Z rings and constricted the liposomes without any accessory proteins. We proposed that the force for constriction was generated by a conformational change from straight to curved pfs. Evidence supporting this mechanism came from switching the membrane tether to the opposite side of the pf. These switched-tether pfs assembled “inside-out” Z rings, and squeezed the liposomes from the outside, as expected for the bending model. We propose three steps for the full process of cytokinesis: (a) pf bending generates a constriction force on the inner membrane, but the rigid peptidoglycan wall initially prevents any invagination; (b) downstream proteins associate to the Z ring and remodel the peptidoglycan, permitting it to follow the constricting FtsZ to a diameter of ~250 nm; the final steps of closure of the septum and membrane fusion are achieved by excess membrane synthesis and membrane fluctuations.

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“…Unless indicated otherwise, FP constructs contained the full protein sequence and additional 5-aa linkers at the N (GSTLE) and C (LEGST) termini. This is specifically indicated for the mVenus constructs, as we also tested variations in the linkers, as follows: (1), 3 mVenus 229 was missing the first 2 and last 10 aa of the mVenus sequence but retained the 5-aa linkers, and (2), LE-3 mVenus 229 -LE was truncated by an additional 3 aa in each of the linkers.…”

Section: Resultsmentioning

confidence: 99%

Probing for Binding Regions of the FtsZ Protein Surface through Site-Directed Insertions: Discovery of Fully Functional FtsZ-Fluorescent Proteins

et al. 2017

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FtsZ, a bacterial tubulin homologue, is a cytoskeletal protein that assembles into protofilaments that are one subunit thick. These protofilaments assemble further to form a "Z ring" at the center of prokaryotic cells. The Z ring generates a constriction force on the inner membrane and also serves as a scaffold to recruit cell wall remodeling proteins for complete cell division in vivo. One model of the Z ring proposes that protofilaments associate via lateral bonds to form ribbons; however, lateral bonds are still only hypothetical. To explore potential lateral bonding sites, we probed the surface of Escherichia coli FtsZ by inserting either small peptides or whole fluorescent proteins (FPs). Among the four lateral surfaces on FtsZ protofilaments, we obtained inserts on the front and back surfaces that were functional for cell division. We concluded that these faces are not sites of essential interactions. Inserts at two sites, G124 and R174, located on the left and right surfaces, completely blocked function, and these sites were identified as possible sites for essential lateral interactions. However, the insert at R174 did not interfere with association of protofilaments into sheets and bundles in vitro. Another goal was to find a location within FtsZ that supported insertion of FP reporter proteins while allowing the FtsZ-FPs to function as the sole source of FtsZ. We discovered one internal site, G55-Q56, where several different FPs could be inserted without impairing function. These FtsZ-FPs may provide advances for imaging Z-ring structure by superresolution techniques.IMPORTANCE One model for the Z-ring structure proposes that protofilaments are assembled into ribbons by lateral bonds between FtsZ subunits. Our study excluded the involvement of the front and back faces of the protofilament in essential interactions in vivo but pointed to two potential lateral bond sites, on the right and left sides. We also identified an FtsZ loop where various fluorescent proteins could be inserted without blocking function; these FtsZ-FPs functioned as the sole source of FtsZ. This advance provides improved tools for all fluorescence imaging of the Z ring and may be especially important for superresolution imaging.KEYWORDS green fluorescent protein, mEos, mMaple, superresolution, tubulin, Venus F tsZ (filamentous temperature sensitive Z), a bacterial homologue of tubulin, assembles the cytoskeletal framework of the Z ring, which constricts to divide the cell. FtsZ forms protofilaments (pfs), which are considered the building blocks of the Z ring. The Z ring serves as a scaffold for recruitment of downstream cell division proteins, which are mostly involved in remodeling the peptidoglycan wall, generating the septum and eventually the new poles. FtsZ by itself can generate a constriction force when reconstituted in liposomes and is thought to provide a primary force to constrict

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Defining the rate-limiting processes of bacterial cytokinesis

Cited by 161 publications

References 125 publications

Bacterial Division: FtsZ Treadmills to Build a Beautiful Wall

Bacterial Division: FtsZ Treadmills to Build a Beautiful Wall

FtsZ Constriction Force – Curved Protofilaments Bending Membranes

Probing for Binding Regions of the FtsZ Protein Surface through Site-Directed Insertions: Discovery of Fully Functional FtsZ-Fluorescent Proteins

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