PXD046660 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | SUZ12 immunoprecipitation followed by protein indentification via mass spectrometry (SUZ12 IP-MS) in mESCs of SUZ12 WT, delta exon 4, KO, and KO rescues with SUZ12-L and SUZ12-S |
Description | Suz12 exon 4 encodes 23 amino acids (aa 129–152 in SUZ12-L) that partially overlap with the WD-binding domain 1 (WDB1, 110–145). We reasoned that exon 4 skipping might alter the structure of SUZ12 and, possibly, PRC2 composition. To explore this possibility, we generated ESCs that lack the Suz12 exon 4 via CRISPR-Cas9–induced deletion. In addition, to rule out any biases due to the expression levels and/or SUZ12 epitope masking, we generated Suz12 knockout (KO) ESCs (herein, KO) in which Suz12 expression was subsequently rescued by re-introducing either the Suz12-L or Suz12-S mouse isoform fused to a triple-Flag tag under the regulation of a CAG promoter (KO+L/S; Figures S2H–S2K). We performed SUZ12 immunoprecipitation coupled with mass spectrometry (IP-MS) in the WT and ∆ex4 clones to compare their interactomes and with a flag antibody in the KO and rescue cell lines. As expected, no peptides corresponding to exon 4 were retrieved in ∆ex4 samples, while the rest of the sequence displayed similar coverage. Comparison of interactors in the two conditions revealed that SUZ12 binding to AEBP2 and JARID2 was strongly reduced in ∆ex4 cells with respect to WT cells, whereas SUZ12 binding to most core components or to PRC2.1-specific factors was unchanged or only slightly increased . These observations were confirmed by SUZ12 IP followed by Western blot (WB). Flag IP-MS in rescue cells confirmed that, while the long isoform was able to correctly form comparable amounts of both PRC2.1 and PRC2.2 subtypes, interaction of the SUZ12-S with PRC2.2-specific factors was drastically reduced. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-22 |
AnnouncementXML | Submission_2024-10-22_06:33:51.838.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Niccolò Arecco |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; |
ModificationList | No PTMs are included in the dataset |
Instrument | LTQ Orbitrap Velos Pro |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-11-03 14:05:24 | ID requested | |
1 | 2024-03-18 03:53:47 | announced | |
⏵ 2 | 2024-10-22 06:33:52 | announced | 2024-10-22: Updated project metadata. |
Publication List
10.1016/j.molcel.2024.02.011; |
Arecco N, Mocavini I, Blanco E, Ballar, é C, Libman E, Bonnal S, Irimia M, Di Croce L, Alternative splicing decouples local from global PRC2 activity. Mol Cell, 84(6):1049-1061.e8(2024) [pubmed] |
Keyword List
submitter keyword: Flag-tag, Rescues, KO, IP, PRC2, Interactions,mESCs, ESCs, SUZ12, Polycomb |
Contact List
Niccolò Arecco |
contact affiliation | Centre for Genomic Regulation (CRG), Barcelona, Spain |
contact email | arecco.niccolo@gmail.com |
lab head | |
Niccolò Arecco |
contact affiliation | CRG |
contact email | arecco.niccolo@gmail.com |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD046660
- Label: PRIDE project
- Name: SUZ12 immunoprecipitation followed by protein indentification via mass spectrometry (SUZ12 IP-MS) in mESCs of SUZ12 WT, delta exon 4, KO, and KO rescues with SUZ12-L and SUZ12-S