Replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1

Ölmezer, Gizem; Levikova, Maryna; Klein, Dominique; Falquet, Benoît; Fontana, Gabriele; Ćejka, Petr; Rass, Ulrich

doi:10.1038/ncomms13157

Cited by 32 publications

(63 citation statements)

References 70 publications

(110 reference statements)

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“…For example, Sgs1 is also involved later in the HR pathway, including the dissolution of double Holliday junctions leading to non‐crossovers as well as in the regulation of aberrant HR (Oh et al , ; Daley et al , ). Dna2 is engaged in Okazaki fragment maturation (Bae et al , ), in processing of replication intermediates upon stress (Hu et al , ; Thangavel et al , ; Ölmezer et al , ), and in checkpoint signaling (Kumar & Burgers, ).…”

Section: Introductionmentioning

confidence: 99%

Competing interaction partners modulate the activity of Sgs1 helicase during DNA end resection

et al. 2019

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DNA double-strand break repair by homologous recombination employs long-range resection of the 5 0 DNA ends at the break points. In Saccharomyces cerevisiae, this process can be performed by the RecQ helicase Sgs1 and the helicase-nuclease Dna2. Though functional interplay between them has been shown, it remains unclear whether and how these proteins cooperate on the molecular level. Here, we resolved the dynamics of DNA unwinding by Sgs1 at the single-molecule level and investigated Sgs1 regulation by Dna2, the single-stranded DNA-binding protein RPA, and the Top3-Rmi1 complex. We found that Dna2 modulates the velocity of Sgs1, indicating that during end resection both proteins form a functional complex and couple their activities. Sgs1 drives DNA unwinding and feeds single-stranded DNA to Dna2 for degradation. RPA was found to regulate the processivity and the affinity of Sgs1 to the DNA fork, while Top3-Rmi1 modulated the velocity of Sgs1. We hypothesize that the differential regulation of Sgs1 activity by its protein partners is important to support diverse cellular functions of Sgs1 during the maintenance of genome stability.

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Section: Introductionmentioning

confidence: 99%

Competing interaction partners modulate the activity of Sgs1 helicase during DNA end resection

et al. 2019

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“…Genetic inactivation of either the helicase or nuclease activity of DNA2 in cells from a wide range of organisms, including yeast and humans, induces permanent cell cycle arrest (Budd & Campbell, 1997;Budd et al, 2000;Lee et al, 2000;Zheng et al, 2008;Duxin et al, 2009Duxin et al, , 2012. In yeast, the cell death has been ascribed to the role of yeast Dna2 in processing DNA replication intermediates Kang et al, 2000Kang et al, , 2010Olmezer et al, 2016). In humans, DNA2depleted cells undergo arrest during late-S/G2 phase of the cell cycle (Duxin et al, 2009(Duxin et al, , 2012.…”

Section: Introductionmentioning

confidence: 99%

hDNA 2 nuclease/helicase promotes centromeric DNA replication and genome stability

Liu

Jin

et al. 2018

The EMBO Journal

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DNA2 is a nuclease/helicase that is involved in Okazaki fragment maturation, replication fork processing, and end resection of DNA double‐strand breaks. Similar such helicase activity for resolving secondary structures and structure‐specific nuclease activity are needed during DNA replication to process the chromosome‐specific higher order repeat units present in the centromeres of human chromosomes. Here, we show that DNA2 binds preferentially to centromeric DNA. The nuclease and helicase activities of DNA2 are both essential for resolution of DNA structural obstacles to facilitate DNA replication fork movement. Loss of DNA2‐mediated clean‐up mechanisms impairs centromeric DNA replication and CENP‐A deposition, leading to activation of the ATR DNA damage checkpoints at centromeric DNA regions and late‐S/G2 cell cycle arrest. Cells that escape arrest show impaired metaphase plate formation and abnormal chromosomal segregation. Furthermore, the DNA2 inhibitor C5 mimics DNA2 knockout and synergistically kills cancer cells when combined with an ATR inhibitor. These findings provide mechanistic insights into how DNA2 supports replication of centromeric DNA and give further insights into new therapeutic strategies.

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“…Genetic and cell biological experiments offer few insights into the function of the DNA2 helicase. These experiments are limited by the fact that DNA2 and its nuclease activity are required for viability in yeast cells, while both helicase and nuclease activities are essential in human cells, making phenotypic analysis associated with DNA2 defects challenging (Formosa and Nittis 1999; Duxin et al 2012;Wanrooij and Burgers 2015;Olmezer et al 2016).…”

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The motor activity of DNA2 functions as an ssDNA translocase to promote DNA end resection

2017

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DNA2 nuclease-helicase functions in DNA replication and recombination. This requires the nuclease of DNA2, while, in contrast, the role of the helicase activity has been unclear. We now show that the motor activity of both recombinant yeast and human DNA2 promotes efficient degradation of long stretches of ssDNA, particularly in the presence of the replication protein A. This degradation is further stimulated by a direct interaction with a cognate RecQ family helicase, which functions with DNA2 in DNA end resection to initiate homologous recombination. Consequently, helicase-deficient yeast cells display reduced resection speed of HO-induced DNA double-strand breaks. These results support a model of DNA2 and the RecQ family helicase partner forming a bidirectional motor machine, where the RecQ family helicase is the lead helicase, and the motor of DNA2 functions as a ssDNA translocase to promote degradation of 5'-terminated DNA.

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Replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1

Cited by 32 publications

References 70 publications

Competing interaction partners modulate the activity of Sgs1 helicase during DNA end resection

Competing interaction partners modulate the activity of Sgs1 helicase during DNA end resection

hDNA 2 nuclease/helicase promotes centromeric DNA replication and genome stability

The motor activity of DNA2 functions as an ssDNA translocase to promote DNA end resection

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