Similar Datasets
Project description:Next Generation Sequencing Facilitates Quantitative Analysis of control, SETD6 Knock-out, SETD3 knock-out and SETD6+SETD3 knock-out HeLa cells Transcriptomes
| PRJNA819059 | ENA
Project description:The minichromosome maintenance complex (MCM) DNA helicase is an important replicative factor during DNA replication. The proper chromatin loading of MCM is a key step to ensure replication initiation during G1/S phase. Because replication initiation is regulated by multiple biological cues, additional changes to MCM may provide deeper understanding towards this event. Here, we uncover that the histidine methyltransferase SETD3 promotes DNA replication in an enzymatic activity dependent manner. Nascent-strand sequencing (NS-seq) shows that SETD3 regulates replication initiation, as depletion of SETD3 attenuates early replication origins firing. Mechanistically, biochemical experiments reveal that SETD3 binds MCM mainly during G1/S phase, which is required for CDT1-mediated chromatin loading of MCM. The MCM loading relies on the histidine-459 methylation (H459me) on MCM7 that is catalyzed by SETD3. Impairment of H459 methylation attenuates DNA synthesis and chromatin loading of MCM. Furthermore, we show that CDK2 phosphorylates SETD3 at Serine-21 during the G1/S phase, which is required for DNA replication and cell cycle progression. These findings demonstrate a novel mechanism by which SETD3 methylates MCM to regulate replication initiation.
2024-03-12 | GSE243937 | GEO
Project description:With self-renewal and pluripotency features, embryonic stem cells (ESCs) provide an invaluable tool to investigate early cell fate decisions. Pluripotency exit and lineage commitment depend on precise regulation of gene expression that requires coordination between transcription (TF) and chromatin factors in response to various signaling pathways. SET domain-containing 3 (SETD3) is a methyltransferase that can modify histones in the nucleus and actin in the cytoplasm. Through an shRNA screen, we previously identified SETD3 as an important factor in meso/endodermal lineage commitment of mouse ESCs (mESC). In this study, we identified SETD3-dependent transcriptomic changes during endoderm differentiation of (mESCs) using time-course RNA-seq analysis. We found that SETD3 is involved in timely activation of the endoderm-related gene network. Canonical Wnt signaling pathway was one of the markedly altered signaling pathways in the absence of SETD3. The assessment of Wnt transcriptional activity revealed a significant reduction in setd3∆ mESCs coincident with a decrease in nuclear pool of the key TF β-catenin level, though no change was observed in its mRNA or total protein level. Furthermore, proximity ligation assay found an interaction between SETD3 and β-catenin. We were able to rescue the differentiation defect by stably re-expressing SETD3 or activating the canonical Wnt signaling pathway by changing mESC culture conditions. Our results suggest that alterations in canonical Wnt pathway activity and subcellular localization of β-catenin might contribute to the endoderm differentiation defect of setd3∆ mESCs.
2025-03-13 | GSE242033 | GEO
Project description:ACTB is a cytoskeletal protein involved in intracellular trafficking. In recent years, it has become evident that, in addition to its established roles in these compartments, ACTB also participates in the regulation of transcription. However, the molecular mechanisms underlying this function remain poorly understood. The methyltransferase SETD3 has previously been shown to methylate ACTB at H73, thereby regulating ACTB polymerization and smooth muscle contraction. Here, we show that the genomic distribution of ACTB is SETD3-dependent and that this regulation modulates the transcription of genes involved in cell adhesion and mRNA translation in colorectal cancer cells. Proteomic analyses reveal that ACTB and SETD3 interact with multiple large protein complexes, including complexes associated with transcriptional regulation. Specifically, we demonstrate that SETD3-mediated ACTB methylation is required for the co-localization of SMARCA4, a subunit of the SWI/SNF BAF complex, at specific genomic loci. Genomic analyses further show that this co-localization enables the coordinated occupancy of SMARCA4 and H73-methylated ACTB at genes involved in cell adhesion and mRNA translation. Finally, phenotypic assays confirm these regulatory effects. Together, these findings uncover a new mechanistic layer of selective transcriptional regulation mediated by an ACTB–SETD3–SMARCA4 axis in colorectal cancer cells.
2026-01-29 | GSE308375 | GEO
Project description:we employed a TurboID approach that utilized a modified promiscuous biotin ligase BirA for proximity labeling by fusing SETD3 with this enzyme, By introducing biotin, cellular fractions of SETD3-TurboID-transfected cells, both cytosolic and chromatin fractions, underwent immunoprecipitation, followed by MS analysis to identify SETD3-interacting proteins. We conducted two experiments, first time we tested four samples, the RAW data of cyto_Vec and chr_Vec represented cells experssing emptry vector soluble fraction and chromatin fraction respectively. the RAW data of cyto_SET and chr_SET represented cells experssing SETD3-TurboID soluble fraction and chromatin fraction respectively. And PSMs.xlsx described the analysis of mass spectrum data. Second time we tested three samples. The RAW data of 21010704_DHG_VEC, 21010704_DHG_WT and 21010704_DHG_YA represented cells experssing emptry vector, SETD3-WT-TurboID or SETD3-Y313A-TurboID respectively. 21010704_DHG_Protein-compare.xlsx described the proteins identification of mass spectrum data. 21010704_DHG_Methyl-H.xlsx listed the histidine methylation identification of mass spectrum data.
2024-04-16 | PXD050556 | Pride
Project description:we employed a TurboID approach that utilized a modified promiscuous biotin ligase BirA for proximity labeling by fusing SETD3 with this enzyme, By introducing biotin, cellular fractions of SETD3-TurboID-transfected cells, both cytosolic and chromatin fractions, underwent immunoprecipitation, followed by MS analysis to identify SETD3-interacting proteins. We conducted two experiments, first time we tested four samples, the RAW data of cyto_Vec and chr_Vec represented cells experssing emptry vector soluble fraction and chromatin fraction respectively. the RAW data of cyto_SET and chr_SET represented cells experssing SETD3-TurboID soluble fraction and chromatin fraction respectively. And PSMs.xlsx described the analysis of mass spectrum data. Second time we tested three samples. The RAW data of 21010704_DHG_VEC, 21010704_DHG_WT and 21010704_DHG_YA represented cells experssing emptry vector, SETD3-WT-TurboID or SETD3-Y313A-TurboID respectively. 21010704_DHG_Protein-compare.xlsx described the proteins identification of mass spectrum data. 21010704_DHG_Methyl-H.xlsx listed the histidine methylation identification of mass spectrum data.
2024-03-12 | PXD046472 | Pride
Project description:ACTB is a cytoskeletal protein involved in intracellular trafficking. In recent years, it has become evident that, in addition to its established roles in these compartments, ACTB also participates in the regulation of transcription. However, the molecular mechanisms underlying this function remain poorly understood. The methyltransferase SETD3 has previously been shown to methylate ACTB at H73, thereby regulating ACTB polymerization and smooth muscle contraction. Here, we show that the genomic distribution of ACTB is SETD3-dependent and that this regulation modulates the transcription of genes involved in cell adhesion and mRNA translation in colorectal cancer cells. Proteomic analyses reveal that ACTB and SETD3 interact with multiple large protein complexes, including complexes associated with transcriptional regulation. Specifically, we demonstrate that SETD3-mediated ACTB methylation is required for the co-localization of SMARCA4, a subunit of the SWI/SNF BAF complex, at specific genomic loci. Genomic analyses further show that this co-localization enables the coordinated occupancy of SMARCA4 and H73-methylated ACTB at genes involved in cell adhesion and mRNA translation. Finally, phenotypic assays confirm these regulatory effects. Together, these findings uncover a new mechanistic layer of selective transcriptional regulation mediated by an ACTB–SETD3–SMARCA4 axis in colorectal cancer cells.
2026-01-29 | GSE308376 | GEO