<scp>DNA</scp>
            damage kinase signaling: checkpoint and repair at 30 years

Lanz, Michael; Dibitetto, Diego; Smolka, Marcus B.

doi:10.15252/embj.2019101801

Cited by 222 publications

(199 citation statements)

References 266 publications

(337 reference statements)

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“…Accumulated evidence over the past 30 years has identified Mec1 as a central player in the preservation of genome stability, particularly through the regulation of distinct steps of HR such as DNA end resection (Lanz et al, 2019). However, the precise mechanisms and targets regulated by Mec1 signaling for ensuring proper HR control remain incompletely understood, representing a major knowledge gap in our understanding of genome maintenance mechanisms.…”

Section: Discussionmentioning

confidence: 99%

“…In its canonical mode of action, Mec1 activates the downstream kinase Rad53 to mediate a DNA damage checkpoint response that arrests the cell cycle and reshapes the transcriptional and replication programs (Desany, Alcasabas, Bachant, & Elledge, 1998; Huang, Zhou, & Elledge, 1998; Lanz, Dibitetto, & Smolka, 2019; Seeber, Dion, & Gasser, 2013). Mec1 also phosphorylates a range of other targets to mediate checkpoint-independent responses (BastosdeOliveira et al, 2015; Lanz et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…Mec1-dependent stabilization of Rad9 at DNA lesions antagonizes resection (Clerici, Trovesi, Galbiati, Lucchini, & Longhese, 2014; Liu et al, 2017). Mec1 also phosphorylates the DNA repair scaffold Slx4, which counteracts Rad9 recruitment and therefore alleviates the block in resection (Dibitetto et al, 2015; Lanz et al, 2019; Liu et al, 2017; Ohouo, Bastos De Oliveira, Liu, Ma, & Smolka, 2013). Therefore, Mec1 balances anti- and pro-resection outcomes for proper resection control.…”

Section: Introductionmentioning

confidence: 99%

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Phosphoproteomics Reveals a Distinct Mode of Mec1/ATR Signaling in Response to DNA End Hyper-Resection

Sanford

Faça

Vega

et al. 2020

Preprint

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1 2 The Mec1/ATR kinase is crucial for genome maintenance in response to a range of 3 genotoxic insults, although how it promotes context-dependent signaling and DNA 4 repair remains elusive. Here we uncovered a specialized mode of Mec1/ATR signaling 5 triggered by the extensive nucleolytic processing (resection) of DNA ends. Cells lacking 6 RAD9, a checkpoint activator and an inhibitor of resection, exhibit a selective increase 7 in Mec1-dependent phosphorylation of proteins associated with single strand DNA 8 transactions, including the ssDNA binding protein Rfa2, the translocase/ubiquitin ligase 9 Uls1 and the HR-regulatory Sgs1-Top3-Rmi1 (STR) complex. Extensive Mec1-10

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Phosphoproteomics Reveals a Distinct Mode of Mec1/ATR Signaling in Response to DNA End Hyper-Resection

Sanford

Faça

Vega

et al. 2020

Preprint

Self Cite

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“…The DNA damage checkpoint is crucial to protect genome integrity 1,2 . However, the early embryos of many species sacrifice this safeguard to allow for rapid cleavage divisions that are required for speedy development.…”

Section: Main Textmentioning

confidence: 99%

Excess histone H3 is a Chk1 inhibitor that controls embryonic cell cycle progression

Shindo

Amodeo

2020

Preprint

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The early embryos of many species undergo a switch from rapid, reductive cleavage divisions to slower, cell fate-specific division patterns at the Mid-Blastula Transition (MBT). The maternally loaded histone pool is used to measure the increasing ratio of nuclei to cytoplasm (N/C ratio) to control MBT onset, but the molecular mechanism of how histones regulate the cell cycle has remained elusive. Here, we show that excess histone H3 inhibits the DNA damage checkpoint kinase Chk1 to promote cell cycle progression in the Drosophila embryo. We find that excess H3tail that cannot be incorporated into chromatin is sufficient to shorten the embryonic cell cycle and reduce the activity of Chk1 in vitro and in vivo. Removal of the Chk1 phosphosite in H3 abolishes its ability to regulate the cell cycle. Mathematical modeling quantitatively supports a mechanism where changes in H3 nuclear concentrations over the final cell cycles leading up to the MBT regulate Chk1-dependent cell cycle slowing. We provide a novel mechanism for Chk1 regulation by H3, which is crucial for proper cell cycle remodeling during early embryogenesis.

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“…Single-stranded DNA (ssDNA) generated in the wake of uncoupled CMG plays an important physiological role by providing a platform for the activation of the apical checkpoint kinase, Mec1-Ddc2/ATR-ATRIP (budding yeast/vertebrates) 26,27 . Downstream activation of the effector kinase, Rad53/CHK1, elicits a host of responses aimed at preserving genome integrity during replication stress, including cell cycle arrest, increased dNTP levels, nuclease inhibition, and inhibition of origin firing [28][29][30] .…”

Section: Introductionmentioning

confidence: 99%

Rad53 controls DNA unwinding after helicase-polymerase uncoupling at DNA replication forks

Devbhandari

Remus

2019

Preprint

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The coordination of DNA unwinding and synthesis at replication forks promotes efficient and faithful replication of chromosomal DNA. Using the reconstituted budding yeast DNA replication system, we demonstrate that Pol e variants harboring catalytic point mutations in the Pol2 polymerase domain, contrary to Pol2 polymerase domain deletions, inhibit DNA synthesis at replication forks by displacing Pol d from PCNA/primer-template junctions, causing excessive DNA unwinding by the replicative DNA helicase, CMG, uncoupled from DNA synthesis. Mutations that suppress the inhibition of Pol d by Pol e restore viability in Pol2 polymerase point mutant cells. We also observe uninterrupted DNA unwinding at replication forks upon dNTP depletion or chemical inhibition of DNA polymerases, demonstrating that leading strand synthesis is not tightly coupled to DNA unwinding by CMG. Importantly, the Rad53 kinase controls excessive DNA unwinding at replication forks by limiting CMG helicase activity, suggesting a mechanism for fork-stabilization by the replication checkpoint. RESULTS Regulated replication of nucleosome-free plasmid templates in vitroPol2 polymerase point mutations, unlike Pol2 catalytic domain deletions, are lethal 8 . Yet, fork rates in the presence of a catalytic mutant of Pol e (Pol e D640A ) or Pol e lacking the catalytic domain (Pol e Dcat ) appeared to be similar in vitro 13,37 . Therefore, to investigate potential differences between these Pol e variants, we decided to characterize them side-by-side using the reconstituted origin-dependent budding yeast DNA replication system 13,37 . We had previously demonstrated that nucleosomes limit Okazaki fragment length in vitro by restricting strand-displacement synthesis by Pol d 37 . However, using higher salt and lower Pol d concentrations we observe Okazaki fragments with a physiological length distribution also on naked DNA templates ( Figure S1).The modified conditions support efficient maturation of nascent strands by Cdc9 and Fen1 and normal replication of plasmid DNA templates (Figures 1A+B). In the presence of Top2, the two predominant ligation products were full-length or close to full-length linears (ssL), and covalently closed circles (ccc) (Figures 1A).The latter are an expected product of termination, while the former may result from fork stalling upon convergence 38 . Linear full-length or close to full-length daughter strands were also generated in the presence of Top1. As expected, due to the inability of Top1 to decatenate double-stranded DNA molecules, circular daughter strands remain topologically linked (cat). The ratio of circular to linear daughter strands was reduced in the presence of Top1 compared to Top2, indicating that Top2 has a greater proficiency in promoting replication termination. On average, we observe a termination efficiency of ~30 % in the presence of Top1 and ~45 % in the presence of Top2 within 45 minutes after origin firing on 4.8 kbp plasmid templates (Figure S2A).In the absence of topoisomerase, replication forks stalled ~ ...

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DNA damage kinase signaling: checkpoint and repair at 30 years

Cited by 222 publications

References 266 publications

Phosphoproteomics Reveals a Distinct Mode of Mec1/ATR Signaling in Response to DNA End Hyper-Resection

Phosphoproteomics Reveals a Distinct Mode of Mec1/ATR Signaling in Response to DNA End Hyper-Resection

Excess histone H3 is a Chk1 inhibitor that controls embryonic cell cycle progression

Rad53 controls DNA unwinding after helicase-polymerase uncoupling at DNA replication forks

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