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Mol Cell. 2015 Nov 5;60(3):374-84. doi: 10.1016/j.molcel.2015.09.009. Epub 2015 Oct 17.

Anionic Phospholipids Stabilize RecA Filament Bundles in Escherichia coli.

Author information

1
Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA.
2
Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Theoretical Chemistry Institute, University of Wisconsin-Madison, Madison, WI 53706, USA.
3
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
4
Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: weibel@biochem.wisc.edu.

Abstract

We characterize the interaction of RecA with membranes in vivo and in vitro and demonstrate that RecA binds tightly to the anionic phospholipids cardiolipin (CL) and phosphatidylglycerol (PG). Using computational models, we identify two regions of RecA that interact with PG and CL: (1) the N-terminal helix and (2) loop L2. Mutating these regions decreased the affinity of RecA to PG and CL in vitro. Using 3D super-resolution microscopy, we demonstrate that depleting Escherichia coli PG and CL altered the localization of RecA foci and hindered the formation of RecA filament bundles. Consequently, E. coli cells lacking aPLs fail to initiate a robust SOS response after DNA damage, indicating that the membrane acts as a scaffold for nucleating the formation of RecA filament bundles and plays an important role in the SOS response.

PMID:
26481664
PMCID:
PMC4771187
DOI:
10.1016/j.molcel.2015.09.009
[Indexed for MEDLINE]
Free PMC Article

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