Visualizing the dynamics of DNA replication and repair at the single-molecule level

Berger, Scott E.; Chistol, Gheorghe

doi:10.1016/bs.mcb.2023.07.001

Cited by 2 publications

(1 citation statement)

References 76 publications

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“…GINS-depleted NPE was supplemented with GINS AF647 (GINS labeled with Alexa Fluor 647) and Fen1 mKikGR (a marker of DNA synthesis 24 ). The replication reaction was monitored using a Total Internal Reflection Fluorescence (TIRF) microscope 25 .…”

Section: A Single-molecule Assay To Visualize Replication Initiation ...mentioning

confidence: 99%

Single-Molecule Imaging Reveals the Mechanism of Bidirectional Replication Initiation in Metazoa

Terui,

Berger,

Sambel

et al. 2024

Preprint

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Metazoan genomes are copied bidirectionally from thousands of replication origins. Replication initiation entails the assembly and activation of two CMG (Cdc45*Mcm2-7*GINS) helicases at each origin. This requires several firing factors (including TopBP1, RecQL4, DONSON) whose exact roles remain unclear. How two helicases are correctly assembled and activated at every single origin is a long-standing question. By visualizing the recruitment of GINS, Cdc45, TopBP1, RecQL4, and DONSON in real time, we uncovered a surprisingly dynamic picture of initiation. Firing factors transiently bind origins but do not travel with replisomes. Two Cdc45 simultaneously arrive at each origin and two GINS are recruited together, even though neither protein can dimerize. The synchronized delivery of two GINS is mediated by DONSON, which acts as a dimerization scaffold. We show that RecQL4 promotes DONSON dissociation and facilitates helicase activation. The high fidelity of bidirectional origin firing can be explained by a Hopfield-style kinetic proofreading mechanism.

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Section: A Single-molecule Assay To Visualize Replication Initiation ...mentioning

confidence: 99%

Single-Molecule Imaging Reveals the Mechanism of Bidirectional Replication Initiation in Metazoa

Terui,

Berger,

Sambel

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Single-molecule fluorescence imaging of DNA maintenance protein binding dynamics and activities on extended DNA

Irvin,

Wang

2024

Current Opinion in Structural Biology

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Visualizing the dynamics of DNA replication and repair at the single-molecule level

Cited by 2 publications

References 76 publications

Single-Molecule Imaging Reveals the Mechanism of Bidirectional Replication Initiation in Metazoa

Single-Molecule Imaging Reveals the Mechanism of Bidirectional Replication Initiation in Metazoa

Single-molecule fluorescence imaging of DNA maintenance protein binding dynamics and activities on extended DNA

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