Proteomics

Dataset Information

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PALB2 interactome in asynchronous human cells

ABSTRACT: Identification of interacting partners of the Partner and Localizer of BRCA2 (PALB2), essential regulator of DNA repair by homologous recombination. Interacting partners of PALB2 were purified by GFP pull down from asynchronous HEK293 Flp-In T-REx cells, exogenously expressing Flag-EGFP tagged PALB2 at a similar level than the endogenous PALB2 level. Before GFP pull down, whole cell protein extracts were pre-cleared on IgG agarose beads to decrease binding of non-specific proteins during GFP pull down. GFP pull down were performed using GFP-Trap agarose beads (Chromotek) and washed at low salt concentration (150mM NaCl), to maintain interactions of low affinity binding protein partners. As a negative control, Flag-EGFP interacting proteins were purified, following the same protocol as described for the purification of Flag-EGFP PALB2 interacting proteins. Experiments were performed in triplicate.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Marjorie Fournier  

LAB HEAD: Fumiko Esashi

PROVIDER: PXD006391 | Pride | 2017-07-18

REPOSITORIES: Pride

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Dataset's files

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Action DRS
HEK293_Flag-EGFPPALB2_Repl1.msf Msf
HEK293_Flag-EGFPPALB2_Repl1.raw Raw
HEK293_Flag-EGFPPALB2_Repl2.msf Msf
HEK293_Flag-EGFPPALB2_Repl2.raw Raw
HEK293_Flag-EGFPPALB2_Repl3.msf Msf
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Publications

MRG15-mediated tethering of PALB2 to unperturbed chromatin protects active genes from genotoxic stress.

Bleuyard Jean-Yves JY   Fournier Marjorie M   Nakato Ryuichiro R   Couturier Anthony M AM   Katou Yuki Y   Ralf Christine C   Hester Svenja S SS   Dominguez Daniel D   Rhodes Daniela D   Humphrey Timothy C TC   Shirahige Katsuhiko K   Esashi Fumiko F  

Proceedings of the National Academy of Sciences of the United States of America 20170703 29

The partner and localiser of BRCA2 (PALB2) plays important roles in the maintenance of genome integrity and protection against cancer. Although PALB2 is commonly described as a repair factor recruited to sites of DNA breaks, recent studies provide evidence that PALB2 also associates with unperturbed chromatin. Here, we investigated the previously poorly described role of chromatin-associated PALB2 in undamaged cells. We found that PALB2 associates with active genes through its major binding part  ...[more]

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