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Biochemical Assays for Analyzing Activities of ATP-dependent Chromatin Remodeling Enzymes

Biochemical Assays for Analyzing Activities of ATP-dependent Chromatin Remodeling Enzymes

Published on: October 25, 2014

ParA ATPases can move and position DNA and subcellular structures.

Florian Szardenings¹, David Guymer, Kenn Gerdes

¹Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, NE2 4HH, Newcastle, United Kingdom.

Current Opinion in Microbiology

|October 4, 2011

Summary

Partitioning proteins (ParA) segregate prokaryotic plasmids via an unknown mechanism, potentially involving filaments. ParA homologues also organize other cellular structures, suggesting a conserved organizational role.

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Area of Science:

Microbiology
Molecular Biology
Cell Biology

Background:

Prokaryotic chromosomes and plasmids utilize partitioning (par) loci for active segregation.
The ParA protein family is known to arrange plasmids into regular arrays over the nucleoid, but the mechanism remains unclear.
ParA is hypothesized to move genetic material via a pulling mechanism, with filament formation being a key question.

Purpose of the Study:

To compare the 'diffusion-ratchet' model with a proposed filament model for ParA-mediated plasmid movement.
To investigate the in vivo presence of ParA filaments, which are only observed in vitro at high concentrations for ParA of P1.
To explore the broader role of ParA homologues in organizing subcellular structures.

Main Methods:

Comparative analysis of proposed mechanisms for ParA-mediated plasmid segregation.
Review of existing in vitro and in vivo data regarding ParA filament formation.
Examination of ParA homologue functions in the arrangement of carboxysomes and chemotaxis receptors.

Main Results:

ParA of P1 forms filaments in vitro only at very high concentrations, and these structures have not been observed in vivo.
A 'diffusion-ratchet' mechanism has been proposed for ParA of P1 plasmid movement.
ParA homologues are found to organize subcellular structures like carboxysomes and chemotaxis receptors into regular arrays, similar to plasmid arrays.

Conclusions:

The precise in vivo mechanism of ParA-mediated plasmid segregation, whether via filaments or another process, requires further elucidation.
The discovery of ParA homologues organizing diverse subcellular structures suggests a conserved and fundamental role in cellular organization beyond plasmid partitioning.