Novel insights into RAD51 activity and regulation during homologous recombination and DNA replication

Godin, Stephen K.; Sullivan, Meghan R.; Bernstein, Kara A.

doi:10.1139/bcb-2016-0012

Cited by 98 publications

(93 citation statements)

References 136 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…S2F), while total ATM expression remained unchanged. In addition, Rad51, a single-stranded DNA-binding protein responsible for HR repair (21,22), showed modest increases in CTCF depleted cells. In contrast, the ATR kinase was only minimally affected by the knockdown (Fig.…”

Section: Ctcf Prevents Genomic Instabilitymentioning

confidence: 99%

CTCF prevents genomic instability by promoting homologous recombination-directed DNA double-strand break repair

Lang

Zheng

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

CTCF is an essential epigenetic regulator mediating chromatin insulation, long-range regulatory interactions, and the organization of large topological domains in the nucleus. Phenotypes of CTCF haploinsufficient mutations in humans, knockout in mice, and depletion in cells are often consistent with impaired genome stability, but a role of CTCF in genome maintenance has not been fully investigated. Here, we report that CTCF maintains genome stability, is recruited to sites of DNA damage, and promotes homologous recombination repair of DNA double-strand breaks (DSBs). CTCF depletion increased chromosomal instability, marked by chromosome breakage and end fusions, elevated genotoxic stress-induced genomic DNA fragmentation, and activated the ataxia telangiectasia mutated (ATM) kinase. We show that CTCF could be recruited to drug-induced 53BP1 foci and known fragile sites, as well as to I-SceI endonuclease-induced DSBs. Laser irradiation analysis revealed that this recruitment depends on ATM, Nijmegen breakage syndrome (NBS), and the zinc finger DNA-binding domain of CTCF. We demonstrate that CTCF knockdown impaired homologous recombination (HR) repair of DSBs. Consistent with this, CTCF knockdown reduced the formation of γ-radiation-induced Rad51 foci, as well as the recruitment of Rad51 to laser-irradiated sites of DNA lesions and to I-SceI-induced DSBs. We further show that CTCF is associated with DNA HR repair factors MDC1 and AGO2, and directly interacts with Rad51 via its C terminus. These analyses establish a direct, functional role of CTCF in DNA repair and provide a potential link between genome organization and genome stability.CTCF | chromatin | genome stability | DNA repair | homologous recombination

show abstract

Section: Ctcf Prevents Genomic Instabilitymentioning

confidence: 99%

CTCF prevents genomic instability by promoting homologous recombination-directed DNA double-strand break repair

Lang

Zheng

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…How these modifications affect the dynamics of nucleoprotein filament assembly and stability remains to be determined. More importantly, the RAD51-DNA interactions are controlled by the recombination mediator BRCA2, by RAD51 paralogs and Shu complex (Zelensky et al, 2014, Prakash et al, 2015, Godin et al, 2016), by antirecombinogenic DNA helicases (Daley et al, 2014), and by the heteroduplex rejection machinery (Spies and Fishel, 2015). These modulators of RAD51 activity or their individual RAD51-interacting domains can be incorporated into the FRET-based, single-molecule experiments described here.…”

Section: Discussionmentioning

confidence: 99%

Observation and Analysis of RAD51 Nucleation Dynamics at Single-Monomer Resolution

Subramanyam

Kinz-Thompson

Gonzalez

et al. 2018

Methods in Enzymology

View full text Add to dashboard Cite

Human RAD51 promotes accurate DNA repair by homologous recombination and is involved in protection and repair of damaged DNA replication forks. The active species of RAD51 and related recombinases in all organisms is a nucleoprotein filament assembled on single-stranded DNA (ssDNA). The formation of a nucleoprotein filament competent for the recombination reaction, or for DNA replication support, is a delicate and strictly regulated process, which occurs through filament nucleation followed by filament extension. The rates of these two phases of filament formation define the capacity of RAD51 to compete with the ssDNA-binding protein RPA, as well as the lengths of the resulting filament segments. Single-molecule approaches can provide a wealth of quantitative information on the kinetics of RAD51 nucleoprotein filament assembly, internal dynamics, and disassembly. In this chapter, we describe how to setup a single-molecule total internal reflection fluorescence microscopy (TIRFM) experiment to monitor the initial steps of RAD51 nucleoprotein filament formation in real time and at single-monomer resolution. This approach is based on the unique, stretched-ssDNA conformation within the recombinase nucleoprotein filament and follows the efficiency of Förster resonance energy transfer (EFRET) between two DNA-conjugated fluorophores. We will discuss the practical aspects of the experimental setup, extraction of the FRET trajectories, and how to analyze and interpret the data to obtain information on RAD51 nucleation kinetics, the mechanism of nucleation, and the oligomeric species involved in filament formation.

show abstract

“…Rad51 overexpression leads to the accumulation of abnormal karyotypes and gene mutations, which are closely related to carcinogenesis and drug resistance [16]. Studies have shown that, such as breast cancer, pancreatic cancer and other tumors overexpressed Rad51 protein, positively correlated to tumor histological grade and stage [17,18]. In addition, domestic and foreign researchers also found that gene polymorphism of Rad51 recognized as the risk factor of CRC carcinogenesis.…”

Section: Discussionmentioning

confidence: 99%

O-Glcnac Glycosylation of Rad51 Plays an Important Role in Promoting Colorectal Cancer Cell Invasion

Li¹,

Cong²,

Yang³

et al. 2018

J Biomol Res Ther

View full text Add to dashboard Cite

Novel insights into RAD51 activity and regulation during homologous recombination and DNA replication

Cited by 98 publications

References 136 publications

CTCF prevents genomic instability by promoting homologous recombination-directed DNA double-strand break repair

CTCF prevents genomic instability by promoting homologous recombination-directed DNA double-strand break repair

Observation and Analysis of RAD51 Nucleation Dynamics at Single-Monomer Resolution

O-Glcnac Glycosylation of Rad51 Plays an Important Role in Promoting Colorectal Cancer Cell Invasion

Contact Info

Product

Resources

About