Genome-wide mapping sites of H2B-SUMO chromatin binding in WT and set1 mutant
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PROVIDER: PRJNA638143 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:To map H2B-SUMO binding sites on a genome-wide scale, we carried out ChDIP experiment combined with sequencing assay
2020-10-27 | GSE152034 | GEO
Project description:Genome-wide mapping sites of Set3-TAP chromatin binding in WT and H2B 4KA mutant
| PRJNA638144 | ENA
Project description:Histone H3K4 tri-methylation (H3K4me3) catalyzed by Set1/COMPASS, is a prominent epigenetic mark found in promoter-proximal regions of actively transcribed genes. H3K4me3 relies on prior monoubiquitination at the histone H2B (H2Bub) by Rad6 and Bre1. Swd2/Cps35, a Set1/COMPASS component, has been proposed as a key player in facilitating H2Bub-dependent H3K4me3. However, a more comprehensive investigation regarding the relationship among Rad6, Swd2 and Set1 is required to further understand the mechanisms and functions of the H3K4 methylation. We investigated the genome-wide occupancy patterns of Rad6, Swd2 and Set1 under various genetic conditions, aiming to clarify the roles of Set1 and Rad6 for occupancy of Swd2. Swd2 peaks appear on both 5’region and 3’region of genes, which are overlapped with its tightly bound two complexes, Set1 and CPF (Cleavage and Polyadenylation Factor), respectively. In the absence of Rad6/H2Bub, Set1 predominantly localized to the 5ʹ region of genes, while Swd2 lost all the chromatin binding. However, in the absence of Set1, Swd2 occupancy near the 5’region was impared and rather increased in the 3’ region. This study highlights that catalytic activity of Rad6 is essential for all the ways of Swd2’s binding to the transcribed genes and Set1 redistributes the Swd2 to 5’region for accomplishments of H3K4me3 in the genome-wide level.
2024-04-21 | GSE234564 | GEO
Project description:SUMOylation is a reversible post-translational modification regulating all nuclear processes. Identification of SUMOylation sites by mass spectrometry has been hampered by bulky tryptic fragments, which thus far necessitated the use of mutated SUMO. Here, we present a dataset generated through a SUMO-specific protease-based methodology which circumvents this problem, dubbed Protease-Reliant Identification of SUMO Modification (PRISM). PRISM allows for detection of SUMOylated proteins as well as identification of specific sites of SUMOylation while using wild-type SUMO. The method is generic and could be widely applied to study lysine post-translational modifications. PRISM was employed in combination with high-resolution mass spectrometry to identify SUMOylation sites from HeLa cells under standard growth conditions and in response to heat shock. 751 wild-type SUMOylation sites on endogenous proteins were identified, including 200 dynamic SUMO sites in response to heat shock. Thus, we have developed the first method capable of quantitatively studying wild-type mammalian SUMO at the site-specific and system-wide level.
2017-03-28 | MSV000080714 | MassIVE
Project description:The stimulation of trimethylation of histone H3 lysine 4 (H3K4) by H2B monoubiquitination (H2Bub) has been widely studied with multiple mechanisms proposed for this form of histone crosstalk. Cps35/Swd2 within COMPASS is considered to bridge these processes. However, a truncated form of Set1 (762-Set1) is reported to function in H3K4 trimethylation without interacting with Cps35/Swd2, and such crosstalk is attributed to the n-SET domain of Set1 and its interaction with the Cps40/Spp1 subunit of COMPASS. Here, we use biochemical, structural, in vivo, and ChIP-seq approaches to demonstrate that Cps40/Spp1 and the n-SET domain of Set1 are required for the stability of Set1 and not the crosstalk. Furthermore, the apparent wild-type levels of H3K4 trimethylation (H3K4me3) in the 762-Set1 strain is due to rogue methylase activity of this mutant resulting in the mislocalization of H3K4me3 from the promoter-proximal regions to gene bodies and intergenic regions. We have also performed detailed screens and identified yeast strains lacking H2Bub, but containing intact H2Bub enzymes, that have normal levels of H3K4me3, suggesting that ubiquitination may not directly stimulate COMPASS, but rather works in a context of the PAF and Rad6/Bre1 complexes. Our study demonstrates that the ubiquitination machinery and Cps35/Swd2 function to focus COMPASS’ H3K4me3 activity at promoter-proximal regions in a context dependent manner. ChIP-Seq for H3K4ME3 in S. cerevisie wild-type strains and strains expressing a truncated form of Set1: aa762-1080 Set1. H3K4ME3 ChIP-Seq was also compared for wild-type, leo1 knockout, and chd1 knockout strains
2014-01-09 | E-GEOD-53241 | biostudies-arrayexpress
Project description:SUMOylation is a reversible post-translational modification regulating all nuclear processes. Identification of SUMOylation sites by mass spectrometry has been hampered by bulky tryptic fragments, which thus far necessitated the use of mutated SUMO. Here, we present a dataset generated through a SUMO-specific protease-based methodology which circumvents this problem, dubbed Protease-Reliant Identification of SUMO Modification (PRISM). PRISM allows for detection of SUMOylated proteins as well as identification of specific sites of SUMOylation while using wild-type SUMO. The method is generic and could be widely applied to study lysine post-translational modifications. PRISM was employed in combination with high-resolution mass spectrometry to identify SUMOylation sites from HeLa cells under standard growth conditions and in response to heat shock. 751 wild-type SUMOylation sites on endogenous proteins were identified, including 200 dynamic SUMO sites in response to heat shock. Thus, we have developed the first method capable of quantitatively studying wild-type mammalian SUMO at the site-specific and system-wide level.
2015-06-15 | PXD001798 | Pride
Project description:H3K4me3 is catalyzed by the Set1/MLL family of methyltransferases, whose function in catalyzing H3K4me3 is unique. Impaired function of Set1/MLL family members can lead to many abnormalities, such as bone and nerve defects, leukemia, and even death. Although the Set1 family plays an important regulatory role in various biological processes, it is still unclear how the Set1 protein itself is regulated and how protein levels are maintained. Due to the numerous homologues, complex composition, and high molecular weight of Set1 in higher organisms, especially humans, related research is greatly limited. In brewing yeast, Set1 is the only methyltransferase that catalyzes H3K4me3 and is highly conserved between species. Therefore, yeast is an ideal model for studying the functions and mechanisms of the Set1 family. In addition, Set1 protein plays an important role in regulating gene transcription, promoting telomere silencing, and maintaining cell lifespan. The Set1 family also plays an important regulatory role in the occurrence and development of various cancers.
2023-10-24 | PXD042010 | Pride
Project description:We developed a novel method for the identification of SUMO sites by expression of His-tagged SUMO mutants and affinity purification of SUMOylated proteins, followed by trypsin digestion and immunocapture of peptides containing diglycine signature tags. Lab Head: Dr Pascale Cossart, pascale.cossart@pasteur.fr Institut Pasteur Unité des Interactions Bactéries-Cellules, Inserm U604, INRA USC2020 25 rue du Dr. Roux 75015 Paris France
2014-07-29 | PXD000459 | Pride
Project description:In Penicillium oxalicum, histone H2B (PDE_01489) was one of the interacting proteins with the methyltransferase LaeA. It suggests that histone H2B may be a direct target of LaeA/Lae1. To verify whether histone H2B is one of the LaeA targets, an in vitro catalytic experiment was performed using Homo sapiens recombinant histone H2B (HsH2B, expressed in Escherichia coli, without histone modification), SAM as substrates, and LaeA. The products were analyzed by LC-MS/MS for histone modifications. Results showed that the mono and di-methylation of lysine 108 and the mono-methylation of lysine 116 were detected.
2021-07-08 | PXD027171 | iProX
Project description:Histone H2B^3-32, H2B K->G, H2B^3-37, and H2B^30-37 mutations
| PRJNA94007 | ENA