Proteomics

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Ubiquitome analysis of UHRF1 and UHRF2-deficient mESCs

ABSTRACT: The E3 ligase UHRF1 is an essential epigenetic cofactor for DNMT1 dependent maintenance DNA methylation, which provides a binding platform for DNMT1 by both cooperative binding of histones and hemi-methylated DNA as well as by ubiquitinating histone H3. Here, we conduct a comprehensive screen to identify novel ubiquitination targets of UHRF1 and its paralogue UHRF2 by comparing the ubiquitome of wildtype (wt), UHRF1- and UHRF2-deficient mouse embryonic stem cells. With an antibody-dependent enrichment of ubiquitin remnant motif-containing peptides followed by isobaric-labeling based quantitative mass spectrometry, we find both known and novel E3 ligase substrates of UHRF1 involved in a variety of biological processes such as RNA processing, DNA methylation and DNA damage repair.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Stem Cell, Cell Culture

SUBMITTER: Elisabeth Karg  

LAB HEAD: Heinrich Leonhardt

PROVIDER: PXD006593 | Pride | 2017-10-24

REPOSITORIES: pride

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Dataset's files

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Action DRS
MS_rawfile_details.xlsx Xlsx
Ref1028_ES_20170220_U1r1.raw Raw
Ref1028_ES_20170220_U1r2.raw Raw
Ref1028_ES_20170220_U1r3.raw Raw
Ref1028_ES_20170220_U1r4.raw Raw
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Publications

Ubiquitome Analysis Reveals PCNA-Associated Factor 15 (PAF15) as a Specific Ubiquitination Target of UHRF1 in Embryonic Stem Cells.

Karg Elisabeth E   Smets Martha M   Ryan Joel J   Forné Ignasi I   Qin Weihua W   Mulholland Christopher B CB   Kalideris Georgia G   Imhof Axel A   Bultmann Sebastian S   Leonhardt Heinrich H  

Journal of molecular biology 20171018 24

Ubiquitination is a multifunctional posttranslational modification controlling the activity, subcellular localization and stability of proteins. The E3 ubiquitin ligase ubiquitin-like PHD and RING finger domain-containing protein 1 (UHRF1) is an essential epigenetic factor that recognizes repressive histone marks as well as hemi-methylated DNA and recruits DNA methyltransferase 1. To explore enzymatic functions of UHRF1 beyond epigenetic regulation, we conducted a comprehensive screen in mouse e  ...[more]

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