Proteomics

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Chromatin proteome of S-phase in IgG- and DONSON-depleted egg extract

ABSTRACT: The aim of the project was to analyse the role of DONSON during DNA replication reaction in Xenopus laevis egg extract. To test what is the impact of DONSON on replication we decided to remove DONSON from egg extract by immunodepletion and analyse its impact on DNA replication reaction. In the parallel experiments we have shown that without DONSON egg extract cannot synthesise nascent DNA. Here we wanted to examine the change of replicating chromatin proteome in absence of DONSON. Interphase Xenopus laevis egg extract was immunodepleted with control nonspecific IgG antibodies beads or DONSON-antibodies. The efficiency of immunodepletion was shown to be over 95%. DNA replication was set up in both IgG- and DONSON-depleted egg extracts and chromatin isolated in the middle of S-phase, when replication forks where abundant in chromatin fraction in IgG-depleted extract.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Xenopus Laevis (african Clawed Frog)

TISSUE(S): Cell Culture, Oocyte

SUBMITTER: Aga Gambus  

LAB HEAD: Dr Aga Gambus

PROVIDER: PXD042435 | Pride | 2023-10-24

REPOSITORIES: Pride

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MSB61644ABand_01.raw Raw
MSB61644ABand_01.raw_20230522_Byonic.mzid.mzid.gz Mzid
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Publications

DONSON facilitates Cdc45 and GINS chromatin association and is essential for DNA replication initiation.

Kingsley Georgia G   Skagia Aggeliki A   Passaretti Paolo P   Fernandez-Cuesta Cyntia C   Reynolds-Winczura Alicja A   Koscielniak Kinga K   Gambus Agnieszka A  

Nucleic acids research 20231001 18

Faithful cell division is the basis for the propagation of life and DNA replication must be precisely regulated. DNA replication stress is a prominent endogenous source of genome instability that not only leads to ageing, but also neuropathology and cancer development in humans. Specifically, the issues of how vertebrate cells select and activate origins of replication are of importance as, for example, insufficient origin firing leads to genomic instability and mutations in replication initiati  ...[more]