Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Loss of KNTC1 Triggers a 53BP1 and USP28 Dependent Cell Cycle Arrest Without Prolonged Mitotic Delays

ABSTRACT: Accurate and efficient mitosis is the hallmark of animal cycling cells. While the spindle assembly checkpoint (SAC) ensures mitotic fidelity by delaying mitotic progression, a SAC-independent surveillance program is known to monitor the duration – without affecting the progression – of mitosis. This pathway, mediated by 53BP1, USP28, and p53, induces post-mitotic arrest in daughter cells following mitoses that exceed a defined temporal threshold. To uncover additional inputs into this pathway, we performed genome-wide CRISPR-Cas9 screens in wild-type, p53-/-, and USP28-/- human cells. Among genes selectively required in wild-type cells, we identified two components of the RZZ kinetochore complex—KNTC1 (ROD) and ZWILCH—as high-confidence hits. Remarkably, depleting KNTC1, which impaired SAC maintenance, triggered a robust 53BP1, USP28, and p53-dependent arrest without the characteristic mitotic delay. KNTC1-deficent cells could arrest following multiple, consecutive divisions with normal durations, and the last mitosis prior to arrest progressed no differently from the previous. Instead, loss of KNTC1 was found to trigger 53BP1-USP28-p53 complex formation soon after mitotic entry, an indication of pathway activation without prolonged mitosis. These findings suggest that the 53BP1–USP28–p53 surveillance pathway can monitor various mitotic anomalies to safeguard cell fitness.

ORGANISM(S): Homo sapiens

PROVIDER: GSE306237 | GEO | 2026/03/10

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

CRL3-GMCL1 mediates 53BP1 degradation, promoting evasion from mitotic surveillance upon paclitaxel treatment

Project description:The Mitotic Surveillance Pathway (MSP) is a critical quality control mechanism that monitors mitosis duration. Prolonged mitosis - caused by genomic defects, spindle errors, or anti-mitotic drugs such as paclitaxel - induces formation of the ternary -mitotic stopwatch- complex composed of 53BP1, USP28, and p53. This complex protects p53 from MDM2-mediated degradation, and its accumulation transmits a memory of the delayed mitosis to the daughter cells, where p53 signaling triggers cell cycle arrest and apoptosis. In Paclitaxel-resistant cancers, cells bypass the MSP, enabling unchecked proliferation and survival, though the underlying mechanisms remain unknown. Here, we identify CRL3GMCL1 as the ubiquitin ligase that targets 53BP1 for degradation during prolonged M phase. In cancers with high GMCL1 levels, the CRL3GMCL1-mediated degradation of 53BP1 prevents the formation of the mitotic stopwatch complex, leading to p53 degradation and sustained proliferation. Loss of GMCL1 stabilizes mitotic 53BP1 levels and restores paclitaxel sensitivity. Our results indicate that combining GMCL1 inhibition with taxane treatment represent a novel strategy to restore taxane sensitivity in cancers. The mass spectrometric data files for the protein complex analysis of FLAG-tagged: (i) wild-type GMCL1 (GMCL1 WT), (ii) a GMCL1 E142K mutant (GMCL1 EK for short), which disrupts its ability to bind with CUL3, and (iii) a GMCL1 BTB/BACK-only mutant, which misses GMCL1's putative C-terminal substrate-binding domain are included in this entry.

2025-03-04 | MSV000097235 | MassIVE

53BP1 integrates DNA repair and p53-dependent cell fate decisions via distinct mechanisms

Project description:The tumor suppressor protein 53BP1, a pivotal regulator of DNA double-strand break (DSB) repair, was first identified as a p53-interacting protein over two decades ago, however its direct contributions to p53-dependent cellular activities remain undefined. Here, we reveal 53BP1 stimulates genome-wide p53-dependent gene transactivation and repression events in response to ionizing radiation (IR) and synthetic p53 activation. 53BP1-dependent p53 modulation requires both auto-oligomerization and tandem-BRCT domain mediated bivalent interactions with p53 and the ubiquitin-specific protease USP28. Loss of these activities results in inefficient p53-dependent cell-cycle checkpoint and exit responses. Furthermore, we demonstrate 53BP1-USP28 cooperation to be essential for normal p53-promoter element interactions and gene transactivation-associated events, yet dispensable for 53BP1-dependent DSB repair regulation. Collectively, our data provides a mechanistic explanation for 53BP1-p53 cooperation in controlling anti-tumorigenic cell fate decisions, and reveal these activities to be distinct and separable from 53BP1’s regulation of DNA double-strand break repair pathway choice.

2016-08-18 | GSE84986 | GEO

High proliferation and delamination during skin epidermal stratification

Project description:The development of complex stratified epithelial barriers in mammals is initiated from single-layered epithelia. How stratification is initiated and fueled are still open questions. Previous studies on skin epidermal stratification suggested a central role for perpendicular/asymmetric cell division orientation of the basal keratinocyte progenitors. Here, we use centrosomes, that organize the mitotic spindle, to test whether cell division orientation and stratification are linked. Genetically ablating centrosomes from the developing epidermis leads to the activation of the p53-, 53BP1- and USP28-dependent mitotic surveillance pathway causing a thinner epidermis and hair follicle arrest. The centrosome/p53-double mutant keratinocyte progenitors significantly alter their division orientation in the later stages without majorly affecting epidermal differentiation. Together with time-lapse imaging and tissue growth dynamics measurements, the data suggest that the first and major phase of epidermal development is boosted by high proliferation rates in both basal and suprabasally-committed keratinocytes as well as cell delamination, whereas the second phase maybe uncoupled from the division orientation of the basal progenitors. The data provide insights for tissue homeostasis and hyperproliferative diseases that may recapitulate developmental programs.

2021-04-02 | GSE161387 | GEO

The dimeric deubiquitinase Usp28 integrates 53bp1 and Myc functions to limit DNA damage

Project description:The oncogenic transcription factor Myc is a pleiotropic regulator of RNA Polymerase II (RNAPII)-dependent transcription, DNA replication and DNA damage response pathways. Myc is stringently regulated by the ubiquitin system - for example, ubiquitination controls recruitment of the elongation factor Paf1c, which is critical for several Myc functions. Curiously, a key Myc-targeting deubiquitinase Usp28 also controls cellular response to DNA damage via the mediator protein 53bp1. Usp28 forms stable dimers, but the biological role of Usp28 dimerization is unknown. We show that dimerization limits Usp28 activity and restricts recruitment of Paf1c by Myc. Expression of monomeric Usp28 leads to ectopic Paf1c recruitment and resolution of transcription-replication conflicts, accelerating DNA synthesis. Strikingly, 53bp1 selectively interacts with and stabilizes dimeric Usp28 - depletion of 53bp1 favors formation of Usp28 monomers deregulating DNA replication. Genotoxic stress disrupts 53bp1-Usp28 complexes, promotes formation of Usp28 monomers and recruitment of Paf1 by Myc. This triggers ectopic DNA synthesis during early response to genotoxins, amplifying DNA damage. We propose that dimerization of Usp28 limits aberrant replication at transcriptionally active chromatin to maintain genome stability.

2023-11-09 | PXD037263 | Pride

Functional analysis of Cdc20 reveals a critical role of CRY box in mitotic checkpoint signaling

Project description:Accurate chromosome segregation is coordinated by the spindle assembly checkpoint (SAC) through its effector the mitotic checkpoint complex (MCC), to inhibit the anaphase-promoting complex or cyclosome (APC/C). Cdc20 is an essential mitotic regulator since it promotes mitotic exit through activating the APC/C and monitors kinetochore-microtubule attachment through activating the SAC. The proper functioning of Cdc20 requires multiple interactions with APC/C and MCC subunits. To functionally assess each of these interactions within cells requires efficient depletion of endogenous Cdc20, which is highly difficult to achieve by RNAi. Here we generated Cdc20 RNAi sensitive cell lines by CRISPR/Cas9 which display a penetrant metaphase arrest phenotype by a single RNAi treatment. In this null background, we accurately measured the contribution of each known motif of Cdc20 on APC/C and SAC activation. The CRY box, a previously identified degron was found to be critical for the SAC by promoting theMCC formation and stabilizing the interaction between the MCC and APC/C. These data reveal additionalregulatory components within the SAC and establish a novel method to interrogate Cdc20 function.

2023-12-06 | PXD045411 | Pride

Mitotic stress is an integral part of the oncogene-induced senescence program that promotes multinucleation and cell cycle arrest

Project description:Oncogene-induced senescence (OIS) is a tumor suppression mechanism that blocks cell proliferation in response to oncogenic signalling. OIS is frequently accompanied by multinucleation; however, the origin of this is unknown. Here we show that multinucleate OIS cells originated mostly from failed mitosis. Prior to senescence, mutant RasV12 activation in primary human fibroblasts compromised mitosis, associated with abnormal expression of mitotic genes that enter M-phase. Simultaneously, RasV12 activation enhanced survival of damaged mitoses, culminating in extended mitotic arrest and aberrant exit from mitosis via mitotic slippage. ERK-dependent transcriptional up-regulation of Mcl1 was responsible for enhanced slippage of cells with mitotic defects and subsequent cell survival. Importantly, mitotic slippage and oncogene signalling synergistically induced senescence and key senescence regulators p21 and p16. We propose that activated Ras induces transcriptional changes that predispose cells undergoing OIS to mitotic stress and multinucleation. We used RNA-seq of IMR90 cells with inducible expression of oncogenic RasV12 that were synchronised in mitosis, to characterise the nature of mitotic defects that lead to multinucleation of oncogene-induced senescent cells

2015-08-13 | E-GEOD-70668 | biostudies-arrayexpress

TDP1 phosphorylation at S61 promotes Top1cc repair via the MUS81-dependent pathway during mitosis

Project description:Topoisomerase 1 (Top1) removes supercoils from DNA during replication and transcription, is critical for mitotic progression to the G1 phase, and ensures DNA topology. Tyrosyl-DNA phosphodiesterase 1 (TDP1) mediates the removal of trapped Top1-DNA covalent complexes (Top1ccs). Here, we identify CDK1-dependent phosphorylation of TDP1 at S61 residue during mitosis. TDP1 defective for S61 phosphorylation (TDP1S61A) is trapped on the mitotic chromosomes, triggering DNA damage and mitotic defects. Moreover, we show that Top1cc repair in mitosis occurs via MUS81-dependent mitotic DNA repair mechanism. Replication stress (RS) induced by Camptothecin (CPT) or Aphidicolin (APH) leads to TDP1S61A enrichment at common fragile sites (CFSs), which over-stimulates MUS81-dependent chromatid breaks, anaphase bridges, and micronuclei, ultimately culminating in 53BP1 nuclear bodies in the G1-phase. Our findings provide a new insight into the cell cycle-dependent regulation of TDP1 dynamics forthe repair of trapped Top1ccs during mitosis that prevents genomic instability following RS.

2024-06-26 | PXD053309 | Pride

GATA2 mitotic bookmarking is required for definitive haematopoiesis [ChIP-seq]

Project description:In mitosis, most transcription factors detach from chromatin, but some are retained and bookmark genomic sites. Mitotic bookmarking has been implicated in lineage inheritance, pluripotency and reprogramming. However, the biological significance of this mechanism in vivo remains unclear. Here, we address mitotic retention of the hemogenic factors GATA2, GFI1B and FOS during haematopoietic specification. We show that GATA2 remains bound to chromatin throughout mitosis, in contrast to GFI1B and FOS, via C-terminal zinc finger-mediated DNA binding. GATA2 bookmarks a subset of its interphase targets that are co-enriched for RUNX1 and other regulators of definitive haematopoiesis. Remarkably, homozygous mice harbouring the cyclin B1 mitosis degradation domain upstream Gata2 partially phenocopy knockout mice. Degradation of GATA2 at mitotic exit abolishes definitive haematopoiesis at aorta-gonad-mesonephros, placenta and foetal liver, but does not impair yolk sac haematopoiesis. Our findings implicate GATA2-mediated mitotic bookmarking as critical for definitive haematopoiesis and highlight a dependency on bookmarkers for lineage commitment.

2023-06-27 | GSE207551 | GEO

GATA2 mitotic bookmarking is required for definitive hematopoiesis [scRNA-seq]

2023-06-27 | GSE221691 | GEO

Greatwall kinase regulates SAC by controlling phosphorylation and activity of MPS1

Project description:The Greatwall/Mastl kinase is an essential gene, which regulates the phosphatase activity that counteracts the phosphorylation of Cdk1 substrates. Although Mastl-/- MEFs can enter mitosis, they are unable to complete mitosis due to chromosome segregation defects. Here, we show that Mastl is essential for robust spindle assembly checkpoint (SAC) maintenance. Mastl-/- MEFs display reduced phosphorylation and kinase activity of MPS1, which causes mislocalization of MPS1, Mad1, and Aurora B from the kinetochore and the inner centromere regions. This results in premature inactivation of SAC signalling and early anaphase onset without correction of chromosome-spindle attachment mistakes. Treatment of the knockout cells with protein phosphatase II inhibitor okadaic acid (OA) rescued decreased MPS1 activity, mislocalization of Aurora B, Mad1, MPS1, and premature mitotic slippage. Our data demonstrates how regulation of PP2A activity by Mastl kinase prevents premature SAC silencing as a result of controlling the phosphorylation status and activity of MPS1.

2018-10-26 | PXD004882 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data