Kinetochores Coordinate Pericentromeric Cohesion and Early DNA Replication by Cdc7-Dbf4 Kinase Recruitment

Natsume, Toyoaki; Müller, Carolin A.; Katou, Yuki; Retkute, Renata; Gierliński, Marek; Araki, Hideki; Blow, J. Julian; Shirahige, Katsuhiko; Nieduszynski, Conrad A.; Tanaka, Tomoyuki

doi:10.1016/j.molcel.2013.05.011

Cited by 143 publications

(220 citation statements)

References 50 publications

(77 reference statements)

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“…1B). Conversely, the Ctf19 kinetochore protein advances pericentromeric origin firing by facilitating the modification of centromeric heterochromatin (Natsume et al 2013). The Fkh1 transcription factor also effects origin firing, delaying some origins and advancing others (Knott et al 2012).…”

Section: Resultsmentioning

confidence: 99%

“…(B) The correlation between the firing time of origins, as determined by the firing-time parameter n from a quantitative analysis of replication kinetics ) and the number of MCM ChIP-seq reads in a 1-kb window around those origins. Blue dots represent origins repressed by telomere proximity (Lian et al 2011), Rpd3 (Knott et al 2009) activity, or Fkh1 (Knott et al 2012); red dots represent centromeric origins and other origins activated by Ctf19 (Natsume et al 2013); green dots represent all other origins. Chromosome 10 origins are indicated in orange; the origins used in Figure 2 and Supplemental Figure S2 are indicated in purple.…”

Section: Resultsmentioning

confidence: 99%

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Replication timing is regulated by the number of MCMs loaded at origins

et al. 2015

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Replication timing is a crucial aspect of genome regulation that is strongly correlated with chromatin structure, gene expression, DNA repair, and genome evolution. Replication timing is determined by the timing of replication origin firing, which involves activation of MCM helicase complexes loaded at replication origins. Nonetheless, how the timing of such origin firing is regulated remains mysterious. Here, we show that the number of MCMs loaded at origins regulates replication timing. We show for the first time in vivo that multiple MCMs are loaded at origins. Because early origins have more MCMs loaded, they are, on average, more likely to fire early in S phase. Our results provide a mechanistic explanation for the observed heterogeneity in origin firing and help to explain how defined replication timing profiles emerge from stochastic origin firing. These results establish a framework in which further mechanistic studies on replication timing, such as the strong effect of heterochromatin, can be pursued.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Replication timing is regulated by the number of MCMs loaded at origins

et al. 2015

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“…The existence of mechanisms that target Scc2/Scc4 to key chromosomal regions, such as pericentromeres, may explain why the modest peak of Scc2/Scc4 in late G 1 suffices in early cell cycle periods. Kinetochores directly mediate Scc2/Scc4 enrichment within pericentromeric chromatin, thereby ensuring the robust cohesion of sister chromatids that is vital for promoting chromosome biorientation, even under conditions in which cohesins are limiting (27,39,40). Perhaps more surprising given a previous report that Scc2 is not essential after S phase in the absence of DNA damage (7) are our observations that robust Scc2/ Scc4 chromatin association is achieved in G 2 /M cells and that post-S phase cell viability is reduced following Scc2 inactivation.…”

Section: Discussionmentioning

confidence: 99%

Cell cycle-specific cleavage of Scc2 regulates its cohesin deposition activity

Woodman

Fara

Dzieciątkowska

et al. 2014

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

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Sister chromatid cohesion (SCC), efficient DNA repair, and the regulation of some metazoan genes require the association of cohesins with chromosomes. Cohesins are deposited by a conserved heterodimeric loading complex composed of the Scc2 and Scc4 proteins in Saccharomyces cerevisiae, but how the Scc2/Scc4 deposition complex regulates the spatiotemporal association of cohesin with chromosomes is not understood. We examined Scc2 chromatin association during the cell division cycle and found that the affinity of Scc2 for chromatin increases biphasically during the cell cycle, increasing first transiently in late G 1 phase and then again later in G 2 /M. Inactivation of Scc2 following DNA replication reduces cellular viability, suggesting that this post S-phase increase in Scc2 chromatin binding affinity is biologically relevant. Interestingly, high and low Scc2 chromatin binding levels correlate strongly with the presence of full-length or amino-terminally cleaved forms of Scc2, respectively, and the appearance of the cleaved Scc2 species is promoted in vitro either by treatment with specific cell cycle-staged cellular extracts or by dephosphorylation. Importantly, Scc2 cleavage eliminates Scc2-Scc4 physical interactions, and an scc2 truncation mutant that mimics in vivo Scc2 cleavage is defective for cohesin deposition. These observations suggest a previously unidentified mechanism for the spatiotemporal regulation of cohesin association with chromosomes through cell cycle regulation of Scc2 cohesin deposition activity by Scc2 dephosphorylation and cleavage.M ultisubunit, ring-shaped cohesin complexes play key roles in chromosome morphogenesis that are required for faithful chromosome transmission to daughter cells. Newly replicated sister chromatids become tethered together by cohesins during S phase, which promotes chromosome biorientation on mitotic spindles (1). Cohesins also mediate efficient DNA double-strand break repair by homologous recombination (2, 3) and the formation or stabilization of chromatin loops that affect various nuclear processes, such as gene expression and Ig gene rearrangements (reviewed in refs. 4 and 5). Altered gene expression resulting from defective cohesinmediated chromatin looping is likely responsible for the pathogenesis of Cornelia de Lange Syndrome (CdLS), a dominantly inherited human developmental disorder (6).Sister chromatid cohesion (Scc) proteins form a heterodimeric cohesin deposition complex, but the complex's activity in deposition is not understood (7). Cohesins copurify with Scc2/Scc4, suggesting that Scc2/Scc4 plays a direct role in deposition (8-11). In the absence of either loader complex subunit, cohesin rings assemble, but fail to be deposited (7,12,13). ATP hydrolysis by cohesin's structural maintenance of chromosome (SMC) subunits is required for cohesin loading, and deposition is inhibited when SMC hinge domains, which mediate Smc1/3 interactions within cohesin, are artificially tethered (8,14,15). Thus, Scc2/ Scc4 may activate cohesin's ATPase activity or f...

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“…However, it does not explain the regulation of the replication timing of individual chromosomal domains. Several recent studies showed that DDK is modulated in a chromosome loci specific form; for example, the Ctf19 kinetochore complex can recruit Dbf4 to the kinetochores in telophase to promote early DNA replication in S-phase (Natsume et al 2013). Then again, origins that fire late during S-phase, e.g.…”

Section: Ddk Dependent Regulation Of Dna Replication In Saccharomycesmentioning

confidence: 99%

Switch on the engine: how the eukaryotic replicative helicase MCM2–7 becomes activated

2014

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A crucial step during eukaryotic initiation of DNA replication is the correct loading and activation of the replicative DNA helicase, which ensures that each replication origin fires only once. Unregulated DNA helicase loading and activation, as it occurs in cancer, can cause severe DNA damage and genomic instability. The essential mini-chromosome maintenance proteins 2-7 (MCM2-7) represent the core of the eukaryotic replicative helicase that is loaded at DNA replication origins during G1-phase of the cell cycle. The MCM2-7 helicase activity, however, is only triggered during S-phase once the holo-helicase Cdc45-MCM2-7-GINS (CMG) has been formed. A large number of factors and several kinases interact and contribute to CMG formation and helicase activation, though the exact mechanisms remain unclear. Crucially, upon DNA damage, this reaction is temporarily halted to ensure genome integrity. Here, we review the current understanding of helicase activation;we focus on protein interactions during CMG formation, discuss structural changes during helicase activation and outline similarities and differences of the prokaryotic and eukaryotic helicase activation process.3

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Kinetochores Coordinate Pericentromeric Cohesion and Early DNA Replication by Cdc7-Dbf4 Kinase Recruitment

Cited by 143 publications

References 50 publications

Replication timing is regulated by the number of MCMs loaded at origins

Replication timing is regulated by the number of MCMs loaded at origins

Cell cycle-specific cleavage of Scc2 regulates its cohesin deposition activity

Switch on the engine: how the eukaryotic replicative helicase MCM2–7 becomes activated

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