Proteomics

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DECODING PROTEIN METHYLATION FUNCTION WITH THERMAL STABILITY ANALYSIS


ABSTRACT: Protein methylation is emerging as an important modification beyond epigenetics. However, systems-wide analyses of protein methylation function lag behind compared to other modifications. Recently, thermal stability analyses have been developed which provide a proxy of a protein functional status. Here, we show that diverse molecular and functional events closely linked to protein methylation can be revealed by the analysis of thermal stability. Using mouse embryonic stem cells (mESC) as a model system, we show that Prmt5 regulates numerous mRNA binding proteins that are enriched in intrinsically disordered regions and involved in liquid-liquid phase separation mechanisms. Our data show for instance that methylation of Prmt5 substrates promotes the formation of stress granules. Moreover, we reveal a novel non-canonical function of Ezh2 in mitotic chromosomes and the organization of the perichromosomal layer and identify Mki67 as a putative novel Ezh2 substrate involved in this process. Our approach provides an opportunity to systematically explore protein methylation function and represents a rich resource for understanding the role of this modification in pluripotency.

INSTRUMENT(S): Q Exactive HF-X, Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver, Stem Cell, Cell Culture, Fibroblast, Kidney

SUBMITTER: Cristina Sayago Ferreira  

LAB HEAD: Javier Munoz

PROVIDER: PXD038939 | Pride | 2023-04-27

REPOSITORIES: Pride

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Publications


Protein methylation is an important modification beyond epigenetics. However, systems analyses of protein methylation lag behind compared to other modifications. Recently, thermal stability analyses have been developed which provide a proxy of a protein functional status. Here, we show that molecular and functional events closely linked to protein methylation can be revealed by the analysis of thermal stability. Using mouse embryonic stem cells as a model, we show that Prmt5 regulates mRNA bindi  ...[more]

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