Proteomics

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RB1 mass spec with or without MYC amplification

ABSTRACT: 293T cells were transfected with Flag-tagged RB1 with or without MYC. After 48 h, the cells were lysed with IP buffer and immunoprecipitated with Flag-conjugated agarose beads (Sigma-Aldrich). The bound proteins were eluted with 2% SDS and digested overnight with sequencing grade modified trypsin (Promega, PRV5111). The obtained peptides were analyzed using a nanoflow EASY-nLC 1200 system (Thermo Fisher Scientific, Odense, Denmark) coupled to an Orbitrap Exploris480 mass spectrometer (Thermo Fisher Scientific, Bremen, Germany). The results were processed with the UniProt human protein database (75,004 entries, download on 07-01-2020) using Protein Discoverer (Version 2.4.1.15, Thermo Fisher Scientific) and Mascot (Version 2.7.0, Matrix Science).

ORGANISM(S): Homo Sapiens

SUBMITTER: Jian Ma  

PROVIDER: PXD037479 | iProX | Mon Oct 17 00:00:00 BST 2022

REPOSITORIES: iProX

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Publications

CDK4/6 inhibitors (CDK4/6i) show anticancer activity in certain human malignancies, such as breast cancer. However, their application to other tumor types and intrinsic resistance mechanisms are still unclear. Here, we demonstrate that MYC amplification confers resistance to CDK4/6i in bladder, prostate and breast cancer cells. Mechanistically, MYC binds to the promoter of the E3 ubiquitin ligase KLHL42 and enhances its transcription, leading to RB1 deficiency by inducing both phosphorylated and  ...[more]

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