PXD046640-1

PXD046640 is an original dataset announced via ProteomeXchange.

Title	Histone post translational modification mass sprectrometry SUZ12 exon 4 cell lines
Description	We investigate the contribution of each SUZ12 isoform on PRC2 activity on chromatin using our different ESC lines. As an additional comparison for ∆ex4 cells (SUZ12-S), we included an ESC clone in which an unsuccessful CRISPR editing event resulted in a mutant allele with nearly constitutive splicing of exon 4 (herein, CSex4) and normal Suz12 expression levels (expressing only SUZ12-L). First, we profiled H3K27 bulk PTMs by WB and observed that, while CSex4 cells showed no major differences to control cells, ∆ex4 cells displayed lower levels of H3K27me2 and -me3, with higher levels of mono-methylation and acetylation. Similar results were obtained when comparing SUZ12-L and SUZ12-S ESC rescue lines. To validate these findings with an antibody-independent technique, and to additionally profile other histone PTMs, we analysed these cells with histone-MS. In line with the previous estimates26, WT cells displayed approximately 85% of H3K27 methylation. This proportion was largely similar in CSex4 cells, while it dropped to ~50% in ∆ex4 cells, with H3K27me2 and -me3 being the most affected modifications. Notably, H3K27me2/3 loss in ∆ex4 cells occurred at histone peptides regardless of their H3K36 methylation status. Moreover, the global H3K36 methylation rates were not affected in either CSex4 or ∆ex4 cells (Figure S4C), and no major changes in methylation or acetylation levels were observed at other residues. Importantly, reintroduction of SUZ12-L in KO cells rescued a higher degree of methylation compared to SUZ12-S. Overall, these results indicate that SUZ12-S alone is unable to maintain global physiological levels of H3K27 methylation; rather SUZ12-L is also necessary for this task.
HostingRepository	PRIDE
AnnounceDate	2024-03-18
AnnouncementXML	Submission_2024-03-18_04:07:39.658.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	monomethylated residue; acetylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-03 03:59:32	ID requested
⏵ 1	2024-03-18 04:07:40	announced

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]

submitter keyword: PTMs, KO, histones, PRC2,mESCs, ESCs, SUZ12, DDA, Polycomb

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

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PRIDE project URI

• PRIDE
  1. PXD046640
     1. Label: PRIDE project
     2. Name: Histone post translational modification mass sprectrometry SUZ12 exon 4 cell lines