Cdh1 Proteomic Analysis - A Synthetic Lethal Interaction between APC/C and Topoisomerase Poisons Uncovered by Proteomic Screens

ABSTRACT: The Anaphase-promoting complex/cyclosome (APC/C) cofactor Cdh1 modulates proliferation and cell cycle exit by targeting multiple cell cycle regulators for ubiquitin-dependent degradation. Lack of Cdh1 results in structural and numerical chromosome aberrations, a hallmark of genomic instability, although the specific targets involved in these defects are not well understood. Here, we use MS-based proteomics to identify substrates of Cdh1 important for cell cycle regulation. To this end, we performed three experiments: 1 - SILAC analysis of Cdh1 deficient asynchronous MEFs (samples were fractionated by means of SDS-PAGE and IEF) 2 - iTRAQ analysis of Cdh1 deficient synchronous MEFs (samples were fractionated by means of IEF) 3 - iTRAQ analysis of brain tissue in a Cdh1 deficient Mouse (samples were fractionated by means of IEF) LC-MS was carried out using a LTQ-Orbitrap Velos as described in Eguren et al. iTRAQ raw data was processed using Proteome Discoverer 1.3 interfaced with Mascot 2.2 SILAC raw data was processed using MaxQuant suite using Andromeda as search engine Further details can be found in Eguren et al.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Fernando Garcia  

PROVIDER: PXD000511 | Pride | 2014-02-20

REPOSITORIES: Pride