Escherichia coli FtsA forms lipid-bound minirings that antagonize lateral interactions between FtsZ protofilaments

Marcin Krupka,Veronica W Rowlett,Kara Schoenemann,Dustin Morado,William Margolin,Jun Liu,Heidi Vitrac

doi:10.1038/ncomms15957

Marcin Krupka, Veronica W Rowlett + Show 5 more

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https://doi.org/10.1038/ncomms15957

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Abstract

Most bacteria divide using a protein machine called the divisome that spans the cytoplasmic membrane. Key divisome proteins on the membrane’s cytoplasmic side include tubulin-like FtsZ, which forms GTP-dependent protofilaments, and actin-like FtsA, which tethers FtsZ to the membrane. Here we present genetic evidence that in Escherichia coli, FtsA antagonizes FtsZ protofilament bundling in vivo. We then show that purified FtsA does not form straight polymers on lipid monolayers as expected, but instead assembles into dodecameric minirings, often in hexameric arrays. When coassembled with FtsZ on lipid monolayers, these FtsA minirings appear to guide FtsZ to form long, often parallel, but unbundled protofilaments, whereas a mutant of FtsZ (FtsZ*) with stronger lateral interactions remains bundled. In contrast, a hypermorphic mutant of FtsA (FtsA*) forms mainly arcs instead of minirings and enhances lateral interactions between FtsZ protofilaments. Based on these results, we propose that FtsA antagonizes lateral interactions between FtsZ protofilaments, and that the oligomeric state of FtsA may influence FtsZ higher-order structure and divisome function.

Highlights

Most bacteria divide using a protein machine called the divisome that spans the cytoplasmic membrane
It is well known that an excess of FtsA inhibits E. coli cell division that can be corrected by a compensatory increase in FtsZ34,35
ATP was added to the mixtures, we found that FtsA minirings assembled without adding exogenous ATP, and addition of more ATP had no reproducible effect on miniring assembly (Fig. 3e,f)

Summary

Introduction

Most bacteria divide using a protein machine called the divisome that spans the cytoplasmic membrane. Key divisome proteins on the membrane’s cytoplasmic side include tubulin-like FtsZ, which forms GTP-dependent protofilaments, and actin-like FtsA, which tethers FtsZ to the membrane. It is well established that FtsZ assembles into dynamic GTP-dependent protofilaments, and single-molecule or polarization microscopy experiments suggest that these protofilaments are loosely arranged around the cell membrane in an B100 nm-wide zone at midcell[9,10,11,12] These patches are dynamic, as they have been shown to move in Bacillus subtilis[7], and the FtsZ and FtsA structures within them are dynamic as well, with rapid subunit turnover as measured by photobleaching experiments[13,14]. Consistent with this, a recent study of the in vitro assembly of FtsA and FtsZ on supported lipid bilayers showed that treadmilling FtsZ polymers were more dynamic when tethered to the bilayers by FtsA than by ZipA, suggesting that FtsA promotes turnover of FtsZ subunits[17]

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