ABSTRACT: Localisation of CpG methylation in yeast expressing murine DNMTS Genomic DNA was purified from a control strain and a strain expressing murine DNMTs, treated with Bisulfite and sequenced on a hiseq 2000
Project description:Effect of induced Methylation on Nucleosome positioning in yeast. Mnase digested DNA from a control strain, a strain expressing the 4 murine DNMTs or a strain expressing catalytically inactive murine DNMTs were extracted and sequenced on a hiseq 2000
Project description:Effect of induced Methylation on Nucleosome positioning in yeast. Mnase digested DNA from a control strain and a strain expressing the 4 murine DNMTs were extracted and sequenced on a hiseq 2000
Project description:Effect of induced Methylation on 3D genome organisation in yeast. Hi-C experiment were performed on yeast expressing or not the 4 murine DNMTs (DNMT1, 3a, 3b and 3L).
Project description:Effect of induced Methylation on Gene expression in yeast. Total RNA from a control strain and a strain expressing the 4 murine DNMTs were extracted and sequenced in a hiseq 4000
Project description:Localisation of CpG methylation in yeast expressing murine DNMTS Genomic DNA was purified from a control strain and a strain expressing murine DNMTs
Project description:DNA methylation is an epigenetic modification associated with transcriptional repression of promoters and is essential for mammalian development. Establishment of DNA methylation is mediated by the de novo DNA methyltransferases DNMT3A and DNMT3B, whereas DNMT1 ensures maintenance of methylation through replication. Absence of these enzymes is lethal, and somatic mutations in these genes have been associated with several human diseases. How genomic DNA methylation patterns are regulated remains poorly understood, as the mechanisms that guide recruitment and activity of DNMTs in vivo are largely unknown. To gain insights into this matter we determined chromosomal binding and site-specific activity of the mammalian de novo DNA methyltransferases DNMT3A and DNMT3B. We show that both enzymes localize to methylated, CpG dense regions in mouse stem cells, yet are excluded from active promoters and enhancers. By specifically measuring sites of de novo methylation, we observe that enzymatic activity reflects chromosomal binding. De novo methylation increases with CpG density, yet is excluded from nucleosomes. Notably, we observed selective binding of DNMT3B to the bodies of transcribed genes, which leads to their preferential methylation. This targeting to transcribed sequences requires SETD2-mediated methylation of lysine 36 on histone H3 and a functional PWWP domain of DNMT3B. Together these findings reveal how sequence and chromatin cues guide de novo methyltransferase activity to ensure methylome integrity. Whole-genome bisulfite sequencing for Dnmt1,3a,3b-triple-KO ES cells expressing DNMT3A2 or DNMT3B1 and for Dnmt1,3a,3b,Setd2-KO ES cells expressing DNMT3B1
Project description:What methylation changes are occurring in different parts of early maturation stage seed largely remains unknown. To uncover the possible role of DNA methylation in different parts of early maturation stage seed, we characterized the methylome of seed coats,cotyledons, and the embryonic seed axis using Illumina sequencing. seed coats, cotyledon, and axis
Project description:Seeds are comprised of three majors parts of distinct parental origin: the seed coat, embryo, and endosperm. The maternally-derived seed coat is important for nurturing and protecting the seeds during development. By contrast, the embryo and the endosperm are derived from a double fertilization event, where one sperm fertilizes the egg to form the diploid zygote and the other sperm fertilizes the central cell to form the triploid endosperm. Each seed parts undergo distinct developmental programs during seed development. What methylation changes occurring in the different seed parts, if any, remains unknown. To uncover the possible role of DNA methylation in different parts of the seed, we characterized the methylome of three major parts of an early maturation stage seed: seed coat, embryonic cotyledons, and embryonic axis using Illumina sequencing. Illumina sequencing of bisulfite-converted genomic DNA from three parts of an mid-maturation (B1) stage seed: seed coat (B1-SC), embryonic cotyledons (B1-COT), and embryonic axis (B1-AX).
Project description:The oomycete pathogen Phytophthora sojae causes root rot of soybean. During infection, the pathogen is thought to deliver dozens, if not hundreds, of effector proteins into the host to manipulate intracellular systems. Although these pathogen proteins often exhibit similar N-terminal delivery domains, the remaining effector region is rarely homologous to known protein domains, making it difficult to predict its biochemical function during infection. As a complement to studies in the natural host, Saccharomyces cerevisiae has been successfully used as a model system to explore the biochemical function of individual pathogen effectors. The presumption is that many effectors target conserved eukaryotic pathways in the host and consequently the expression of effectors in yeast will confer corresponding phenotypes. Indeed, putative effector functions identified using yeast functional genomic approaches have subsequently been validated in the natural host. Over-expression of the Phytophthora sojae effector Avh172 (PsAvh172) inhibits the growth of Saccharomyces cerevisiae, suggesting that the effector targets a biological pathway conserved with plants. In this study, the transcriptomes of yeast expressing PsAvh172 or an empty vector were compared to examine the global transcriptional response, in hopes of discerning the effectors biochemical target.