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Single-Molecule DNA Polymerase Dynamics at a Bacterial Replisome in Live Cells
Yi Liao1, Yilai Li1, Jeremy W Schroeder2
1Department of Chemistry, University of Michigan, Ann Arbor, Michigan.
Biophysical Journal
|December 22, 2016
Summary
Bacillus subtilis uses two DNA polymerases for replication. This study reveals the dynamic behavior of PolC, a key polymerase, showing its constant recruitment and release at the replisome.
Area of Science:
- Microbiology
- Molecular Biology
- Biophysics
Background:
- Bacillus subtilis utilizes a two-DNA polymerase system for chromosomal replication, similar to eukaryotes.
- PolC is one of the two essential replicative DNA polymerases in B. subtilis.
Purpose of the Study:
- To quantitatively image and understand the in vivo behavior of the PolC DNA polymerase at single-molecule resolution.
- To elucidate the dynamics and stoichiometry of PolC within the B. subtilis replisome.
Main Methods:
- Photobleaching-assisted microscopy
- Three-dimensional superresolution imaging
- Single-particle tracking
Main Results:
- Determined the stoichiometry of PolC proteins per cell and per replisome.
- Characterized the diffusion patterns of individual PolC molecules in vivo.
- Quantified the exchange dynamics of PolC during lagging strand synthesis, revealing high dynamism.
Conclusions:
- PolC is a highly dynamic DNA polymerase, continuously recruited to and released from the bacterial replisome.
- This dynamic behavior provides new insights into the organization and function of the replisome in bacterial cells.
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