Dataset Information

Phospho-regulation of Cdc48/p97 cofactor protein Shp1/p47 in budding yeast

ABSTRACT: Shp1/p47 is a cofactor of the Cdc48/p97 complex involved in the energy-dependent segregation of intracellular aggregates and multiprotein complexes. We identified two serine residues (S108 and S315) in the S. Cerevisiae protein Shp1 which increase in phosphorylation upon various cell stresses such as heat shock, TORC1 inhibition, ER stress and oxidative stress. To identify binding partners of phosphorylated vs non-phosphorylated Shp1, we generated endogenous C-terminal 5xFLAG-tagged Shp1 yeast strains either harbouring either WT shp1 or CRISPR-modified S108A + S315A (2SA) shp1. These were cultured with 200 nM rapamycin treatment for 2 h to induce phosphorylation, prior to lysis, FLAG immunoprecipitation, peptide digestion and processing for LC-MS/MS.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture

SUBMITTER: Frederic Lamoliatte

LAB HEAD: Adrien Rousseau

PROVIDER: PXD035819 | Pride | 2023-01-30

REPOSITORIES: Pride

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Multiple phosphorylation of the Cdc48/p97 cofactor protein Shp1/p47 occurs upon cell stress in budding yeast.

Agrotis Alexander A Lamoliatte Frederic F Williams Thomas D TD Black Ailsa A Horberry Rhuari R Rousseau Adrien A

Life science alliance 20230124 4

The homohexameric p97 complex, composed of Cdc48 subunits in yeast, is a crucial component of protein quality control pathways including ER-associated degradation. The complex acts to segregate protein complexes in an ATP-dependent manner, requiring the engagement of cofactor proteins that determine substrate specificity. The function of different Cdc48 cofactors and how they are regulated remains relatively poorly understood. In this study, we assess the phosphorylation of Cdc48 adaptor protein ...[more]

PMID: 36693698

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	20210701_M4_1.raw	Raw
	20210701_M4_2.raw	Raw
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	20210701_WT_1.raw	Raw
	20210701_WT_2.raw	Raw