Spatial and temporal organization of RecA in the Escherichia coli DNA-damage response.

Content Provider	Europe PMC
Author	Ghodke, Harshad Paudel, Bishnu P Lewis, Jacob S Jergic, Slobodan Gopal, Kamya Romero, Zachary J Wood, Elizabeth A Woodgate, Roger Cox, Michael M van Oijen, Antoine M
Editor	Ha, Taekjip Kuriyan, John
Abstract	The RecA protein orchestrates the cellular response to DNA damage via its multiple roles in the bacterial SOS response. Lack of tools that provide unambiguous access to the various RecA states within the cell have prevented understanding of the spatial and temporal changes in RecA structure/function that underlie control of the damage response. Here, we develop a monomeric C-terminal fragment of the λ repressor as a novel fluorescent probe that specifically interacts with RecA filaments on single-stranded DNA (RecA*). Single-molecule imaging techniques in live cells demonstrate that RecA is largely sequestered in storage structures during normal metabolism. Upon DNA damage, the storage structures dissolve and the cytosolic pool of RecA rapidly nucleates to form early SOS-signaling complexes, maturing into DNA-bound RecA bundles at later time points. Both before and after SOS induction, RecA* largely appears at locations distal from replisomes. Upon completion of repair, RecA storage structures reform.
Journal	eLife
Volume Number	8
PubMed Central reference number	PMC6363387
PubMed reference number	30717823
e-ISSN	2050084X
DOI	10.7554/elife.42761
Language	English
Publisher	eLife Sciences Publications, Ltd
Publisher Date	2019-02-05
Access Restriction	Open
Rights License	This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
Subject Keyword	SOS response RecA DNA damage DNA repair intermediates live cell imaging single molecule imaging E. coli
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Medicine Biochemistry, Genetics and Molecular Biology