PXD046783

PXD046783 is an original dataset announced via ProteomeXchange.

Title	Proteomic profiling of Palbociclib arrested cells ± APC/C inactivation.
Description	Cellular proliferation is highly regulated to ensure proper tissue homeostasis and to prevent diseases such as cancer. In actively proliferating cells, entry into S phase of the cell cycle and initiation of DNA replication is promoted by inactivation of the E3 ubiquitin ligase APC/C (anaphase promoting complex/cyclosome), though the critical S phase-promoting substrates have not been fully elucidated. The APC/C is also active in cells that have exited the cell cycle and entered an arrested state, but whether APC/C activity is essential to maintain arrest is unclear. We found that APC/C inactivation is sufficient to bypass cellular arrest induced by the CDK4/6 inhibitor Palbociclib. To identify potential APC/C substrates responsible for driving the escape from cell cycle arrest, we arrested cells using Palbociclib and then induced EMI1, inactivating the APC/C. Samples were collected at 0, 8, 16 and 20h after arrest and analyzed using proteomics. We found that inactivation of the APC/C promotes cell cycle re-entry by inducing RB phosphorylation and E2F-dependent gene transcription. Stabilization of both cyclin A and cyclin B contributes to arrest bypass, but only cyclin A accumulation is absolutely required. Direct expression of APC/C-resistant cyclin A, but not cyclin B, in arrested cells induces S phase entry analogous to APC/C inactivation. Cells bypassing arrest initiate DNA replication with severely reduced origin licensing, at least in part due to premature geminin accumulation. As a result, cells exhibit reduced rates of DNA synthesis, which leads to the accumulation of replication stress and long-term proliferation defects. Our findings suggest that CDK4/6 inhibition in cancers with reduced APC/C activity may be ineffective at promoting cytostatic cell cycle arrest but may nonetheless lead to elevated levels of replication stress that ultimately leads to a more durable cell cycle withdrawal.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_06:50:47.838.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Christine Mills
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-08 05:32:49	ID requested
1	2024-07-18 10:21:35	announced
⏵ 2	2024-10-22 06:50:48	announced	2024-10-22: Updated project metadata.

10.1101/2023.11.09.566394;

Mouery BL, Baker EM, Mills CA, Herring LE, Fleifel D, Cook JG, APC/C prevents non-canonical order of cyclin/CDK activity to maintain CDK4/6 inhibitor-induced arrest. bioRxiv, ():(2023) [pubmed]

submitter keyword: cancer,Palbociclib, cell cycle, APC/C

Jean Cook
contact affiliation	Department of Biochemistry and Biophysics, RNA Discovery Center, and Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill
contact email	jean_cook@med.unc.edu
lab head
Christine Mills
contact affiliation	University of North Carolina - Chapel Hill
contact email	allie.mills@unc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD046783
     1. Label: PRIDE project
     2. Name: Proteomic profiling of Palbociclib arrested cells ± APC/C inactivation.