79 research outputs found
The PIDDosome: centrosome guardian and backup on the DNA damage response
No full textThe PIDDosome is a Caspase-2-activating platform assembling in response to centrosome amplification or genotoxic stress. We have recently shown that both stimuli depend on ANKRD26 (ankyrin repeat domain-containing protein 26)-mediated localization of PIDD1 (p53-inducible protein with death domain) at the centrosome, demonstrating how this organelle can directly influence cell fate
CRISPR/Cas9 ribonucleoprotein-mediated knockin generation in hTERT-RPE1 cells
No full textSummary: hTERT-RPE1 cells are genetically stable near diploid cells widely used to model cell division, DNA repair, or ciliogenesis in a non-transformed context. However, poor transfectability and limited homology-directed repair capacity hamper their amenability to gene editing. Here, we describe a protocol for rapid and efficient generation of diverse homozygous knockins. In contrast to other approaches, this strategy bypasses the need for molecular cloning. Our approach can also be applied to a variety of cell types including cancer and induced pluripotent stem cells (iPSCs)
Centriolar distal appendages activate the centrosome‐PIDDosome‐p53 signalling axis via ANKRD26
No full textCentrosome amplification results into genetic instability and predisposes cells to neoplastic transformation. Supernumerary centrosomes trigger p53 stabilization dependent on the PIDDosome (a multiprotein complex composed by PIDD1, RAIDD and Caspase-2), whose activation results in cleavage of p53's key inhibitor, MDM2. Here, we demonstrate that PIDD1 is recruited to mature centrosomes by the centriolar distal appendage protein ANKRD26. PIDDosome-dependent Caspase-2 activation requires not only PIDD1 centrosomal localization, but also its autoproteolysis. Following cytokinesis failure, supernumerary centrosomes form clusters, which appear to be necessary for PIDDosome activation. In addition, in the context of DNA damage, activation of the complex results from a p53-dependent elevation of PIDD1 levels independently of centrosome amplification. We propose that PIDDosome activation can in both cases be promoted by an ANKRD26-dependent local increase in PIDD1 concentration close to the centrosome. Collectively, these findings provide a paradigm for how centrosomes can contribute to cell fate determination by igniting a signalling cascade
Supplementary Data Figure S26 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S26 showing that removal of mutant TP53 does not impact in vivo tumor growth upon transplantation of WCB139T human colon cancer derived organoids into NSG mice</p
Supplementary Data Figure S9 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S9 showing that complete removal of mutant TP53 in Rael-BL cells does not impair survival and proliferation in culture</p
Supplementary Data Figure S14 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S14 showing that CRISPR-HDR mediated conversion of mutant TP53 into wt TP53 causes a growth disadvantage in human cancer cell lines</p
Supplementary Data Figure S28 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S28 showing that removal of mutant TP53 has no impact on the response of tumors derived from human colon cancer derived organoids to 5-FU in vivo</p
Supplementary Data Figure S8 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S8 showing that complete removal of mutant TP53 in HT29 cells does not impair survival and proliferation in culture</p
Supplementary Data Figure S16 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Figure S16 showing that removal of mutant TP53 does not impair in vivo tumor growth of human cancer lines even when limited numbers of cancer cells are transplanted</p
Supplementary Data Table S4 from Loss-of-Function but Not Gain-of-Function Properties of Mutant TP53 Are Critical for the Proliferation, Survival, and Metastasis of a Broad Range of Cancer Cells
No full textSupplementary Table S4 showing the impact of removal of mutant TP53 in human cancer cell lines in vitro by CRISPR/Cas9 by mining the DepMap database</p
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