TY - JOUR
T1 - Species- and C-terminal linker-dependent variations in the dynamic behavior of FtsZ on membranes in vitro
AU - Sundararajan, Kousik
AU - Vecchiarelli, Anthony
AU - Mizuuchi, Kiyoshi
AU - Goley, Erin D.
N1 - Funding Information:
We would like to thank the members of the Goley lab – Elizabeth Meier, PJ Lariviere, Selam Woldemeskel, Anant Bhargava, Allison Daitch, and Chris Mahone – and the Mizuuchi lab – James Taylor and Michiyo Mizuuchi – for helpful discussions that informed design, optimization and analyses described in this work. We would also like to thank Jie Xiao, Xinxing Yang, and Keir Neuman for their suggestions for quantifying FtsZ dynamics on SLBs. Funding for this work was provided by the NIH through R01GM108640 (to E.D.G.) and the intramural research
Funding Information:
We would like to thank the members of the Goley lab ? Elizabeth Meier, PJ Lariviere, Selam Woldemeskel, Anant Bhargava, Allison Daitch, and Chris Mahone ? and the Mizuuchi lab ? James Taylor and Michiyo Mizuuchi ? for helpful discussions that informed design, optimization and analyses described in this work. We would also like to thank Jie Xiao, Xinxing Yang, and Keir Neuman for their suggestions for quantifying FtsZ dynamics on SLBs. Funding for this work was provided by the NIH through R01GM108640 (to E.D.G.) and the intramural research fund for National Institute of Diabetes and Digestive and Kidney Diseases (to K.M).
Publisher Copyright:
© Published 2018. This article is a U.S. Government work and is in the public domain in the USA.
PY - 2018/10
Y1 - 2018/10
N2 - Bacterial cell division requires the assembly of FtsZ protofilaments into a dynamic structure called the ‘Z-ring’. The Z-ring recruits the division machinery and directs local cell wall remodeling for constriction. The organization and dynamics of protofilaments within the Z-ring coordinate local cell wall synthesis during cell constriction, but their regulation is largely unknown. The disordered C-terminal linker (CTL) region of Caulobacter crescentus FtsZ (CcFtsZ) regulates polymer structure and turnover in solution in vitro, and regulates Z-ring structure and activity of cell wall enzymes in vivo. To investigate the contributions of the CTL to the polymerization properties of FtsZ on its physiological platform, the cell membrane, we reconstituted CcFtsZ polymerization on supported lipid bilayers (SLB) and visualized polymer dynamics and structure using total internal reflection fluorescence microscopy. Unlike Escherichia coli FtsZ protofilaments that organized into large, bundled patterns, CcFtsZ protofilaments assembled into small, dynamic clusters on SLBs. Moreover, CcFtsZ lacking its CTL formed large networks of straight filament bundles that underwent slower turnover than the dynamic clusters of wildtype FtsZ. Our in vitro characterization provides novel insights into species- and CTL-dependent differences between FtsZ assembly properties that are relevant to Z-ring assembly and function on membranes in vivo.
AB - Bacterial cell division requires the assembly of FtsZ protofilaments into a dynamic structure called the ‘Z-ring’. The Z-ring recruits the division machinery and directs local cell wall remodeling for constriction. The organization and dynamics of protofilaments within the Z-ring coordinate local cell wall synthesis during cell constriction, but their regulation is largely unknown. The disordered C-terminal linker (CTL) region of Caulobacter crescentus FtsZ (CcFtsZ) regulates polymer structure and turnover in solution in vitro, and regulates Z-ring structure and activity of cell wall enzymes in vivo. To investigate the contributions of the CTL to the polymerization properties of FtsZ on its physiological platform, the cell membrane, we reconstituted CcFtsZ polymerization on supported lipid bilayers (SLB) and visualized polymer dynamics and structure using total internal reflection fluorescence microscopy. Unlike Escherichia coli FtsZ protofilaments that organized into large, bundled patterns, CcFtsZ protofilaments assembled into small, dynamic clusters on SLBs. Moreover, CcFtsZ lacking its CTL formed large networks of straight filament bundles that underwent slower turnover than the dynamic clusters of wildtype FtsZ. Our in vitro characterization provides novel insights into species- and CTL-dependent differences between FtsZ assembly properties that are relevant to Z-ring assembly and function on membranes in vivo.
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U2 - 10.1111/mmi.14081
DO - 10.1111/mmi.14081
M3 - Article
C2 - 30010220
AN - SCOPUS:85054511060
SN - 0950-382X
VL - 110
SP - 47
EP - 63
JO - Molecular Microbiology
JF - Molecular Microbiology
IS - 1
ER -