Multiple RPAs make WRN syndrome protein a superhelicase

Lee, Mina; Shin, Soochul; Uhm, Heesoo; Hong, Heesun; Kirk, Jaewon; Hyun, Kwang Beom; Kulikowicz, Tomasz; Kim, Jae-Hoon; Ahn, Byungchan; Bohr, Vilhelm A.; Hohng, Sungchul

doi:10.1093/nar/gky272

Cited by 30 publications

(55 citation statements)

References 47 publications

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“…Alternatively, more than one protein unit could bind to form an active unwinding complex. The observed behavior was similar to that seen for BLM (Wang et al , ), WRN (Lee et al , ), and the likely BLM homolog from Arabidopsis thaliana , AtRecQ2 (Klaue et al , ). For AtRecQ2, the transition between unwinding and rezipping most likely involves strand switching (Klaue et al , ).…”

Section: Resultssupporting

confidence: 77%

“…DNA unwinding by Sgs1 was found to be highly dynamic, involving many repetitive unwinding events separated by either rapid DNA rezipping or slower DNA rewinding. Sgs1 shares this highly dynamic activity pattern with other RecQ family helicases from prokaryotes (Harami et al , ; Bagchi et al , ) and eukaryotes (Klaue et al , ; Wang et al , ; Lee et al , ). It is thought that the switching between unwinding and rewinding involves repeated strand switching events to allow direction reversals of the helicase (Klaue et al , ).…”

Section: Discussionmentioning

confidence: 92%

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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: Resultssupporting

confidence: 77%

Section: Discussionmentioning

confidence: 92%

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

et al. 2019

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“…This indicated that a burst was likely initiated by the binding of a single Sgs1 unit (a molecule or a complex), which subsequently originated all events of the burst until the protein finally dissociated. Such behavior is similar to that observed for BLM [29], WRN [30] and the likely BLM-homologue from Arabidopsis thaliana, AtRecQ2 [31]. For the latter it has been shown that the transition between unwinding and rezipping most likely involves strand switching that brings the enzyme in a more loosely bound state since it lacks the DNA junction in its wake [31].…”

Section: Resultssupporting

confidence: 73%

“…DNA unwinding by Sgs1 was found to be highly dynamic, involving many repetitive unwinding events separated by either rapid DNA rezipping or slower DNA rewinding. Sgs1 shares this highly dynamic activity pattern with other RecQ family helicases from prokaryotes [45,46] and eukaryotes [29][30][31]. It is thought that the switching between unwinding and rewinding involves repeated strand switching events to allow direction reversals of the helicase [31].…”

Section: Discussionmentioning

confidence: 99%

Competing interactions modulate the activity of Sgs1 during DNA end resection

Kasaciunaite

Fettes

Levikova

et al. 2019

Preprint

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DNA double-strand break repair by homologous recombination employs long-range resection of the 5' 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 has been shown, it remains unclear whether and how the proteins cooperate on the molecular level. Here, we resolved the dynamics of DNA unwinding by Sgs1 at the single molecule level and investigated its 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 the proteins form a physical complex and couple their activities. Sgs1 unwinds DNA and feeds single-stranded DNA to Dna2 for degradation. RPA is found to regulate the processivity and the affinity of Sgs1 to the DNA fork, while Top3-Rmi1 modulated the velocity of Sgs1. We think that the differential regulation of the Sgs1 activity by its protein partners is important to allow diverse cellular functions of Sgs1 during the maintenance of genome stability.

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“…In homologous recombination WRN interacts with RAD52 and increases its strand annealing activity, whist also modulating WRN helicase activity in a structure dependent manner (13). WRN has been shown to interact via acidic repeats between the helicase and nuclease regions with Replication Protein A (RPA) and this interaction is was found to stimulate WRN helicase activity (14)(15)(16). In telomere metabolism WRN has been found to interact with the shelterin complex components TRF2 and POT1, and this interaction stimulates WRN helicase activity (17,18).…”

Section: Introductionmentioning

confidence: 99%

Crystal Structure of the Werner’s Syndrome Helicase

Newman

Lieb

et al. 2020

Preprint

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Werner syndrome helicase (WRN) plays important roles in DNA repair and the maintenance of genome integrity. Germline mutations in WRN give rise to Werner syndrome, a rare autosomal recessive progeroid syndrome that also features cancer predisposition. Interest in WRN as a therapeutic target has increased massively following the identification of WRN as the top synthetic lethal target for microsatellite instable (MSI) cancers. High throughput screens have identified candidate WRN helicase inhibitors, but the development of potent, selective inhibitors would be significantly enhanced by high-resolution structural information.. In this study we have further characterized the functions of WRN that are required for survival of MSI cancer cells, showing that ATP binding and hydrolysis are required for complementation of siRNA-mediated WRN silencing. A crystal structure of the WRN helicase core at 2.2 Å resolutionfeatures an atypical mode of nucleotide binding with extensive contacts formed by motif VI, which in turn defines the relative positioning of the two RecA like domains. The structure features a novel additional β-hairpin in the second RecA and an unusual helical hairpin in the Zn 2+ binding domain; modelling DNA substrates based on existing RecQ DNA complexes suggests roles for these features in the binding of alternative DNA structures..

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Multiple RPAs make WRN syndrome protein a superhelicase

Cited by 30 publications

References 47 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

Competing interactions modulate the activity of Sgs1 during DNA end resection

Crystal Structure of the Werner’s Syndrome Helicase

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