Project description:RNA-Seq analysis of prostate cancer cell line LNCaP treated with vehicle (C), androgen (R), androgen and IMTPPE (R + IMTPPE), androgen and JJ-(+)-450 (androgen + (-)450), androgen and JJ-(-)450 (androgen + (-)450), androgen and enzalutamide (androgen +Enz). To evaluate if our compounds can inhibit AR function specifically and completely, LNCaP mRNA profiles of cells treated with IMTPPE, (+)-JJ-450 and (-)-JJ-450, comparing to enzalutamide. RNA isolation was performed using RNeasy Mini kit (Qiagen), RNA Sequencing was carried out by Genomics Research Core of University of Pittsburgh using Illumina NextSeq 500 system. The sequence reads that passed FASTQC were analyzed at the transcript level. Each sample was mapped to the Human Ensembl reference genome GRCh38. We definied different expression genes with a fold change ≥2.0 and FDR <0.05. Both (-)-JJ-450 and enzalutamide are very specific to AR, with 56 and 186 DE genes comparing to control samples respectively. IMTPPE and (+)-JJ-450 can inhibit most of the androgen responsive genes, but also some other genes were affected. (-)-JJ-450 is a novel compound inhibts AR function specifically and completely, and it is a potential lead compound for the treatment of CRPC, including those resistant to enzalutamide.
Project description:In flies, the chromosomal kinase JIL-1 is responsible for most interphase H3S10 phosphorylation and has been proposed to protect active chromatin from acquiring heterochromatic marks like H3K9me2 and HP1. Here, we show that JIL-1’s targeting to chromatin depends on a new PWWP domain containing protein JASPer (JIL-1 Anchoring and Stabilizing Protein). The JASPer/JIL-1 (JJ)-complex is the major form of the kinase in vivo and is targeted to active genes and telomeric transposons via binding of the PWWP domain of JASPer to H3K36me3 nucleosomes. Put in place, the complex modulates the transcriptional output. JIL-1 and JJ-complex depletion in cycling cells induce small changes in H3K9me2 distribution at active genes and telomeric transposons. Finally, we identified many new interactors of the endogenous JJ-complex and propose that JIL-1 not only prevents heterochromatinisation, but also coordinates chromatin based regulation in the transcribed part of the genome.
Project description:In flies, the chromosomal kinase JIL-1 is responsible for most interphase H3S10 phosphorylation and has been proposed to protect active chromatin from acquiring heterochromatic marks like H3K9me2 and HP1. Here, we show that JIL-1’s targeting to chromatin depends on a new PWWP domain containing protein JASPer (JIL-1 Anchoring and Stabilizing Protein). The JASPer/JIL-1 (JJ)-complex is the major form of the kinase in vivo and is targeted to active genes and telomeric transposons via binding of the PWWP domain of JASPer to H3K36me3 nucleosomes. Put in place, the complex modulates the transcriptional output. JIL-1 and JJ-complex depletion in cycling cells induce small changes in H3K9me2 distribution at active genes and telomeric transposons. Finally, we identified many new interactors of the endogenous JJ-complex and propose that JIL-1 not only prevents heterochromatinisation, but also coordinates chromatin based regulation in the transcribed part of the genome.
Project description:In flies, the chromosomal kinase JIL-1 is responsible for most interphase H3S10 phosphorylation and has been proposed to protect active chromatin from acquiring heterochromatic marks like H3K9me2 and HP1. Here, we show that JIL-1’s targeting to chromatin depends on a new PWWP domain containing protein JASPer (JIL-1 Anchoring and Stabilizing Protein). The JASPer/JIL-1 (JJ)-complex is the major form of the kinase in vivo and is targeted to active genes and telomeric transposons via binding of the PWWP domain of JASPer to H3K36me3 nucleosomes. Put in place, the complex modulates the transcriptional output. JIL-1 and JJ-complex depletion in cycling cells induce small changes in H3K9me2 distribution at active genes and telomeric transposons. Finally, we identified many new interactors of the endogenous JJ-complex and propose that JIL-1 not only prevents heterochromatinisation, but also coordinates chromatin based regulation in the transcribed part of the genome.