Intrinsic checkpoint deficiency during cell cycle re-entry from quiescence

Matson, Jacob P.; House, Amy M.; Grant, Gavin D.; Wu, Huaitong; Perez, Joanna; Cook, Jeanette Gowen

doi:10.1083/jcb.201902143

Cited by 46 publications

(52 citation statements)

References 69 publications

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“…In this sense, various cell synchronization methods have been criticized for creating an artificial situation [10]. Indeed, recent evidence indicates that even a pause before initiating the cell cycle (i.e., entry to quiescence) will affect how the following S-phase is performed [11]. A prime example of a cell cycle pause is a DNA damage checkpoint.…”

Section: Introductionmentioning

confidence: 99%

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Lafranchi

Müllers

Rutishauser

et al. 2020

Cells

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Cells recovering from the G2/M DNA damage checkpoint rely more on Aurora A-PLK1 signaling than cells progressing through an unperturbed G2 phase, but the reason for this discrepancy is not known. Here, we devised a method based on a FRET reporter for PLK1 activity to sort cells in distinct populations within G2 phase. We employed mass spectroscopy to characterize changes in protein levels through an unperturbed G2 phase and validated that ATAD2 levels decrease in a proteasome-dependent manner. Comparing unperturbed cells with cells recovering from DNA damage, we note that at similar PLK1 activities, recovering cells contain higher levels of Cyclin B1 and increased phosphorylation of CDK1 targets. The increased Cyclin B1 levels are due to continuous Cyclin B1 production during a DNA damage response and are sustained until mitosis. Whereas partial inhibition of PLK1 suppresses mitotic entry more efficiently when cells recover from a checkpoint, partial inhibition of CDK1 suppresses mitotic entry more efficiently in unperturbed cells. Our findings provide a resource for proteome changes during G2 phase, show that the mitotic entry network is rewired during a DNA damage response, and suggest that the bottleneck for mitotic entry shifts from CDK1 to PLK1 after DNA damage.

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Section: Introductionmentioning

confidence: 99%

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Lafranchi

Müllers

Rutishauser

et al. 2020

Cells

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“…As a result, cells are unprotected from premature first S-phase entry, which in turn increases the risk of experiencing genome instability. Moreover, inducing G1 extension by inhibiting Cdk2 activity increases the time for MCM loading, thus, improving the impaired licensing checkpoint [33]. In conclusion, defects in origin licensing can lead to DNA under-replication, which constitutes a major source of genomic instability [29,34].…”

Section: Replication Stress and Under-replicated Dna In Mitosismentioning

confidence: 97%

Working on Genomic Stability: From the S-Phase to Mitosis

Ovejero

Bueno

Sacristán

2020

Genes

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Fidelity in chromosome duplication and segregation is indispensable for maintaining genomic stability and the perpetuation of life. Challenges to genome integrity jeopardize cell survival and are at the root of different types of pathologies, such as cancer. The following three main sources of genomic instability exist: DNA damage, replicative stress, and chromosome segregation defects. In response to these challenges, eukaryotic cells have evolved control mechanisms, also known as checkpoint systems, which sense under-replicated or damaged DNA and activate specialized DNA repair machineries. Cells make use of these checkpoints throughout interphase to shield genome integrity before mitosis. Later on, when the cells enter into mitosis, the spindle assembly checkpoint (SAC) is activated and remains active until the chromosomes are properly attached to the spindle apparatus to ensure an equal segregation among daughter cells. All of these processes are tightly interconnected and under strict regulation in the context of the cell division cycle. The chromosomal instability underlying cancer pathogenesis has recently emerged as a major source for understanding the mitotic processes that helps to safeguard genome integrity. Here, we review the special interconnection between the S-phase and mitosis in the presence of under-replicated DNA regions. Furthermore, we discuss what is known about the DNA damage response activated in mitosis that preserves chromosomal integrity.

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“…Furthermore, cells recovering from quiescence were recently shown to be susceptible to 321 replication stress43 . Stable homologous recombination repair intermediates efficiently sustain 322 ATR/Chk1 signalling, leading to an enhanced checkpoint and eventual cell cycle exit in G2 323 phase 9 .…”

mentioning

confidence: 99%

p53-dependent polyploidisation after DNA damage in G2 phase

Middleton

Suman

O’Toole

et al. 2020

Preprint

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9Cell cycle progression in the presence of damaged DNA can lead to accumulation of mutations 10 and pose a risk for tumour development. In response to DNA damage in G2 phase, human 11 cells can be forced to exit the cell cycle in a p53-p21-and APC/C Cdh1 -dependent manner. Cells 12 that exit the cell cycle in G2 phase become senescent, but it is unclear what determines this 13 commitment and whether other cell fates occur. We find that a subset of immortalised RPE-1 14 cells and primary human fibroblasts spontaneously initiate DNA re-replication several days 15 after forced cell cycle exit in G2 phase. By combining single cell tracking for more than a week 16 with quantitative immunofluorescence, we find that the resulting polyploid cells contain 17 increased levels of damaged DNA and frequently exit the cell cycle again in the next G2 phase. 18Subsequently, these cells either enter senescence or commit to another round of DNA re-19 replication, further increasing the ploidy. At least a subset of the polyploid cells show abnormal 20 centrosome numbers or localisation. In conclusion, cells that are forced to exit the cell cycle in 21 G2 phase face multiple choices that lead to various phenotypes, including propagation of cells 22 with different ploidies. Our findings suggest a mechanism by which p53-positive cells can 23 evade senescence that risks genome integrity. 24 25 Main points 26 -Cell cycle exit from G2 phase does not necessarily lead to senescence 27 28 -Resumption of proliferation after G2 phase cell cycle exit starts with DNA replication 29 30 -Successive cell cycle exits lead to propagation of cells with different ploidies 31 Introduction 35Cell cycle progression in the presence of damaged DNA can lead to propagation of mutations. 36In response to damaged DNA, cells pause the cell cycle, in particular prior to initiating DNA 37 replication or cell division. This pause, termed a checkpoint, is poorly sustained over time. In 38 case of excessive or sustained DNA damage, cells therefore permanently block proliferation, 39 either by regulated cell death, or by entering into senescence. In G2 phase, the road to 40 senescence is initiated by p53-and p21-dependent activation of APC/C Cdh1 1-5 . Activation of 41 APC/C Cdh1 leads to degradation of many key regulators of G2 phase and mitosis, thereby re-42 setting the cell cycle. Whereas we have referred to this phenomenon as cell cycle exit 6 , others 43 described it as cell cycle withdrawal 7 or mitosis skip 8 . Common to all is the description that this 44 is the first irreversible step leading to senescence from G2 phase. While it can be modulated 45 by checkpoint duration 8 , efficiency of DNA repair 9 , and CDK activity in G2 phase 10 , the 46 irreversible step rendering cell cycle exit independent of upstream checkpoint kinase signalling 47 is marked by translocation of Cyclin B1 to the nucleus 6-8 . 48 49 While cell cycle exit occurs rapidly in single cells, establishment of senescence is a gradual 50 process involving large changes in gene expression, c...

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Intrinsic checkpoint deficiency during cell cycle re-entry from quiescence

Abstract: Matson et al. find that human cells re-entering the cell cycle from quiescence have an impaired p53-dependent DNA replication origin licensing checkpoint and slow origin licensing. This combination makes every first S phase underlicensed and hypersensitive to replication stress.

Cited by 46 publications

References 69 publications

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

FRET-Based Sorting of Live Cells Reveals Shifted Balance between PLK1 and CDK1 Activities During Checkpoint Recovery

Working on Genomic Stability: From the S-Phase to Mitosis

p53-dependent polyploidisation after DNA damage in G2 phase

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