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

Author:

Ghodke Harshad12ORCID,Paudel Bishnu P12ORCID,Lewis Jacob S12ORCID,Jergic Slobodan12ORCID,Gopal Kamya3ORCID,Romero Zachary J3,Wood Elizabeth A3,Woodgate Roger4ORCID,Cox Michael M3ORCID,van Oijen Antoine M2ORCID

Affiliation:

1. Molecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, Australia

2. Illawarra Health and Medical Research Institute, Wollongong, Australia

3. Department of Biochemistry, University of Wisconsin-Madison, Madison, United States

4. Laboratory of Genomic Integrity, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, United States

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.

Funder

National Institutes of Health

Australian Research Council

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

Reference101 articles.

1. Human Rad51 protein promotes ATP-dependent homologous pairing and strand transfer reactions in vitro;Baumann;Cell,1996

2. DNA strand exchange proteins: a biochemical and physical comparison;Bianco;Frontiers in Bioscience : A Journal and Virtual Library,1998

3. Quantitation of the Inhibition of Hfr x F- Recombination by the Mutagenesis Complex UmuD'C;Boudsocq;Journal of Molecular Biology,1997

4. RecA Protein Self-assembly Multiple Discrete Aggregation States;Brenner;Journal of Molecular Biology,1988

5. Construction of a Recombinase-deficient Mutant recA Protein That Retains Single-stranded DNA-dependent ATPase Activity;Bryant;The Journal of Biological Chemistry,1988

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3