Dataset Information

Human POT1 Prevents Severe Telomere Damage Instability Induced by Homology Directed DNA Repair

ABSTRACT: The evolutionarily conserved POT1 protein binds the single stranded G-rich telomeric DNA and has been implicated in telomeric DNA maintenance and the suppression of DNA damage checkpoint signaling. Here, we explore human POT1 function through genetics and proteomics discovering that the complete absence of POT1 leads to severe telomere maintenance defects that had not been anticipated from previous depletion studies. We determine the telomeric proteome upon POT1-loss by implementing an improved telomeric chromatin isolation protocol. Using quantitative proteomics by tandem mass tags (TMT) we identified a large set of proteins involved in nucleic acid metabolism that engage with telomeres upon loss of POT1. Inactivation of the homology directed repair machinery suppresses POT1-loss mediated telomeric DNA defects. Our results unravel as major function of human POT1 the suppression of telomere instability induced by homology directed repair.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Manfredo Quadroni

LAB HEAD: Joachim Lingner

PROVIDER: PXD016826 | Pride | 2020-10-21

REPOSITORIES: Pride

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Publications

Human shelterin protein POT1 prevents severe telomere instability induced by homology-directed DNA repair.

Glousker Galina G Briod Anna-Sophia AS Quadroni Manfredo M Lingner Joachim J

The EMBO journal 20201019 23

The evolutionarily conserved POT1 protein binds single-stranded G-rich telomeric DNA and has been implicated in contributing to telomeric DNA maintenance and the suppression of DNA damage checkpoint signaling. Here, we explore human POT1 function through genetics and proteomics, discovering that a complete absence of POT1 leads to severe telomere maintenance defects that had not been anticipated from previous depletion studies in human cells. Conditional deletion of POT1 in HEK293E cells gives r ...[more]

PMID: 33073402

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Project description:Background: Canmei formula (CMF) is a traditional Chinese medicine compound with definite effect on the prevention and treatment of colorectal adenoma (CRA). It can prevent the transformation of intestinal inflammation to cancer. This study explored the mechanism of action of CMF in anti-CRA using multi-omics techniques. Method: Mice were randomly divided into 4 groups: blank group (Control), high-fat diet (HFD) + AOM/DSS colorectal adenoma model (ADH) groups, Canmei formula treatment group (ADH-CMF) and sulfasalazine treatment group (Sul). Except for the blank group, ADH model was established in the other 3 groups by intraperitoneal injection with AOM reagent, and then mice was given 2.5%DSS in free drinking water and high-fat diet. The mice in the blank group and ADH group were intragastrically perfused with normal saline, and the mice in the other 2 groups were treated with corresponding drugs for 20 weeks. During this period, the changes of physical signs of mice in each group were observed. The differentially expressed genes and proteins in the Control group, ADH group and ADH-CMF group were detected by RNA-seq transcriptome sequencing and Tandem Mass Tags (TMT) quantitative proteomics. After the combined analysis and verification. The key targets were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results : A total of 2548 differential genes were obtained by transcriptomics analysis, and 45 differential proteins were obtained by proteomics analysis. The results of proteomics data and experimental verification showed that CMF mainly acted on the Phospholysine Phosphohistidine Inorganic Pyrophosphate Phosphatase (LHPP) target. GO analysis showed that the targets of CMF were involved in the biological processes such as cellular process, metabolic process and biological regulation. KEGG analysis showed that those genes were involved in oxidative phosphorylation, cell senescence, and Metabolic pathways. Studies have shown that Lhpp overexpression of Lhpp impeded Colorectal cancer cell growth and proliferation in vitro, and was associated with a change in PI3K/AKT activity.

Dataset Information

Human POT1 Prevents Severe Telomere Damage Instability Induced by Homology Directed DNA Repair

Publications

Human shelterin protein POT1 prevents severe telomere instability induced by homology-directed DNA repair.

Similar Datasets

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