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Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry
Published on: February 15, 2012
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Dynamics of spore coat morphogenesis in Bacillus subtilis
Peter T McKenney1, Patrick Eichenberger
1New York University, Center for Genomics and Systems Biology, Department of Biology, 12 Waverly Place, 8th floor, New York, NY 10003, USA.
Molecular Microbiology
|December 17, 2011
Summary
Bacillus subtilis spore coat assembly involves diverse protein localization dynamics and layered scaffold formation. This process requires transcriptional regulation and a specific channel for coordinated encasement, revealing forespore contributions to coat morphogenesis.
Area of Science:
- Microbiology
- Cell Biology
- Biochemistry
Background:
- Bacillus subtilis spores possess a protective coat composed of over 70 proteins.
- Previous research primarily focused on the structure of mature spore coats.
Purpose of the Study:
- To investigate the dynamic assembly process (morphogenesis) of the Bacillus subtilis spore coat during sporulation.
- To characterize the localization patterns and temporal dynamics of spore coat proteins.
Main Methods:
- Utilized a library of 41 spore coat proteins fused to green fluorescent protein (GFP).
- Examined protein localization dynamics over the sporulation time-course using live imaging.
- Correlated protein localization kinetics with transcriptional regulators of coat gene expression.
- Investigated the role of specific genes and channels in coat assembly.
Main Results:
- Identified six distinct classes of spore coat protein localization dynamics.
- Demonstrated initial assembly of a multi-layered scaffold at one spore pole.
- Observed spore coat encasement occurring in coordinated waves, driven partly by gene transcription.
- Found the SpoIIIAH-SpoIIQ channel to be essential for coat encasement.
- Showed that deletion of sporulation transcription factors halts encasement.
Conclusions:
- Spore coat morphogenesis is a dynamic, multi-step process with distinct protein localization patterns.
- The assembly involves coordinated waves of encasement, influenced by transcriptional regulation and specific transport channels.
- This study highlights the crucial role of the forespore in contributing to spore coat formation.
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