PXD042953-2

PXD042953 is an original dataset announced via ProteomeXchange.

Title	Histone PTM profiling analysis of EZH2 mutants in isogenic cells
Description	The Enhancer of Zeste 2 Polycomb Repressive Complex 2 Subunit (EZH2) is an essential epigenetic modifier able to methylate lysine 27 on histone H3 (H3K27) to induce chromatin compaction, protein complex recruitment and ultimately transcriptional repression. Hematologic malignancies, including Diffuse Large B cell lymphoma (DLBCL) and Acute myeloid leukemia (AML) have shown a high EZH2-mutation frequency (>20%) associated with poor clinical outcomes. Particularly, two distinct oncogenic mutations, so-called gain-of-function (Y641F and A677G) and loss-of-function (H689A and F667I) are found in the catalytic domain of EZH2. In this study, a comprehensive multi-omics approach was employed to characterize downstream effects of H3K27me3 deposition driven by EZH2 mutations. Human embryonic kidney cells (HEK293T) were transfected to generate three stable EZH2 mutants: EZH2(Y641F), EZH2(A677G), and EZH2(H689A/F667I), which were validated via immunoblotting and DIA-MS-based histone profiling assay. The histone profiling assay demonstrated a significant increase of approximately two-fold in H3.1/H3.3K27me3 for Y641F EZH2 mutant. There was a modest increase in the combinatorial PTMs H3.1/H3.3K27me3K36me1 and a significant depletion in H3.1 and H3.3 K27me2. The most depleted peptide was H3.3K27me2K36me2. For the A677G EZH2 mutant, the assay demonstrated an enrichment on H3.1/H3.3K27me3, combinatorial K27me3K36me1 and a slight increase in K27me1 and K18ac but only on H3.1. The most depleted peptide was H3.3K27me2K36me2. The H689A/F667I cell line has the most alterations in global histone PTMs. The most highly enriched were H3.1/H3.3K36me2, H3.1K9me3 and H3.3K36me1. The most depleted modifications were H3.1K23ac and H3.1K27me3.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:12:04.098.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julian Aldana
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	isopropyl lysyl
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-13 12:47:50	ID requested
1	2023-10-24 13:29:08	announced
⏵ 2	2023-11-14 09:12:05	announced	2023-11-14: Updated project metadata.

Aldana J, Gardner ML, Freitas MA, Integrative Multi-Omics Analysis of Oncogenic EZH2 Mutants: From Epigenetic Reprogramming to Molecular Signatures. Int J Mol Sci, 24(14):(2023) [pubmed]

submitter keyword: lymphoma, epigenetics, leukemia,EZH2

Michael A. Freitas
contact affiliation	Cancer Biology and Genetics, Wexner Medical Center, The Ohio State University, Columbus, OH, USA
contact email	freitas.5@osu.edu
lab head
Julian Aldana
contact affiliation	The Ohio State University
contact email	aldanaaroca.1@osu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042953
     1. Label: PRIDE project
     2. Name: Histone PTM profiling analysis of EZH2 mutants in isogenic cells