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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
Published on: December 17, 2013
Two-step assembly dynamics of the Bacillus subtilis divisome
Pamela Gamba1, Jan-Willem Veening, Nigel J Saunders
1Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Medical School, Framlington Place, Newcastle-upon-Tyne, United Kingdom.
Journal of Bacteriology
|May 12, 2009
Summary
Bacterial cell division involves a two-step assembly of the divisome. Early assembly includes FtsZ, FtsA, ZapA, and EzrA, followed by later recruitment of proteins like GpsB and DivIVA, regulated post-transcriptionally.
Area of Science:
- Microbiology
- Molecular Biology
- Cell Biology
Background:
- Bacterial cell division is orchestrated by the divisome, a complex of proteins.
- Understanding the precise timing and order of divisome assembly is crucial for bacterial growth and reproduction.
Purpose of the Study:
- To investigate the in vivo dynamics and temporal hierarchy of divisome assembly in Bacillus subtilis.
- To elucidate the sequential recruitment of division proteins during cell division.
Main Methods:
- Utilized green fluorescent protein (GFP) fusions to track division protein localization in vivo.
- Employed time-series and time-lapse microscopy in germinating spores and vegetative cells.
- Conducted microarray analysis to assess transcriptional changes.
Main Results:
- Demonstrated a two-step assembly process for the Bacillus subtilis divisome.
- Showed early co-assembly of FtsZ, FtsA, ZapA, and EzrA.
- Identified a delayed recruitment of GpsB, FtsL, DivIB, FtsW, Pbp2B, and DivIVA after a significant cell cycle interval.
- Observed that FtsZ overproduction impacts EzrA but not DivIVA temporal assembly.
- Found no significant transcriptional changes in division genes upon FtsZ manipulation.
Conclusions:
- Provided in vivo evidence for a two-step divisome assembly mechanism in Bacillus subtilis.
- Suggested that FtsZ polymerization is not the sole signal for late divisome protein recruitment.
- Supported the hypothesis that bacterial cell division regulation in B. subtilis occurs primarily at the post-transcriptional level.
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