Project description:B cell differentiation is tightly regulated through coordinated changes in metabolism, division, expression of transcription factors, and epigenetic programming mediated by histone modifying enzymes. In this study, we examined the role of an epigenetic writer, the histone H3K9 mono and dimethyltransferse G9a, in regulating the transcriptional programs during B-cell development and plasma cell (PC) formation. Utilizing a B-cell specific G9afl/flCd19Cre/+ conditional mouse model we performed RNA-seq on naive and activated B cells and plasma cells that formed in response to LPS.
Project description:B cell differentiation is tightly regulated through coordinated changes in metabolism, division, expression of transcription factors, and epigenetic programming mediated by histone modifying enzymes. In this study, we examined the role of an epigenetic writer, the histone H3K9 mono and dimethyltransferse G9a, in regulating the chromatin accessibility programs during B-cell development and plasma cell (PC) formation. Utilizing a B-cell specific G9afl/flCd19Cre/+ conditional mouse model we performed ATAC-seq on naive and activated B cells and plasma cells that formed in response to LPS.
Project description:Proliferating progenitor cells undergo unidirectional changes in competence to give rise to post-mitotic progeny of specialized function. These transitions in cell fate typically involve the dynamic regulation of gene expression programs by histone methyltransferase (HMT) complexes. However, the composition, roles and regulation of these HMT complexes in regulating cell fate decisions in vivo are poorly understood. Using affinity purification and mass spectrometry (AP-MS), we identified the uncharacterized C2H2-like zinc finger (ZF) protein ZNF644, which is putatively causally linked to high-grade (degenerative) myopia, as a novel G9a/GLP-interacting protein and co-regulator of histone methylation. Using zebrafish embryos, we characterized the function of two ZNF644 orthologues, znf644a and znf644b, revealing critical neural- specific roles in regulating G9a/H3K9me2-mediated gene silencing in the retina. In addition, by virtue of additional non-overlapping requirements for znf644a and znf644b during retinal differentiation, we gained further insights regarding additional facets of retinal differentiation regulated by the G9a-ZNF644 physical association, such as maintaining retinal cell viability and in transitioning proliferating progenitor cells towards differentiation. Collectively, our data points to the G9a-ZNF644 physical interaction as a critical neural progenitor-specific co-factor of G9a/H3K9me2-mediated gene silencing during neuronal differentiation.
Project description:Posttranslational modifications of histone N-terminal tails influence the status of chromatin and eventually control the transcriptional outcome of a particular gene. As a histone H3K9 methyltransferase (HMTase) in higher eukaryotes, G9a-mediated transcriptional repression is the major epigenetic silencing machinery. UHRF1 (ubiquitin-like with PHD and ring finger domains I) binds to hemi-methylated DNA and plays essential role in maintenance of DNA methylation by recruiting DNMT1. Here, we provide evidence that UHRF1 is transcriptionally downregulated by H3K9 HMTase G9a. We found that increased expression of G9a along with transcription factor YY1 specifically represses UHRF1 transcription. We uncovered showed that G9a regulates UHRF1-mediated H3K23 ubiquitylation and proper DNA replication maintenance by FACS analysis and propose that H3K9 HMTase G9a is a specific epigenetic regulator of UHRF1.
Project description:As methyltransferase G9a has been implicated in immune cell differentiation we have assessed the role of G9a enzymatic inhibitors (UNC0642) in the differentiation of human naïve T-cells under different stimulation conditions.
Project description:As methyltransferase G9a has been implicated in immune cell differentiation we have assessed the role of G9a enzymatic inhibitors (UNC0642) in the differentiation of human naïve T-cells under different stimulation conditions.