Genome-wide maps of chromatin state (heterochromatin H3K9me2 or centromeric histone H3 variant CENP-A/Cnp1) in the meiotic progeny from the crossings of mhf2+ (cen1_inactive) × mhf2+ (cen1_active) and mhf2? (cen1_inactive) × mhf2+ (cen1_active)

We report the high-throughput profiling of histone modification (H3K9me2) or histone variant CNEP-A/Cnp1 in fission yeast Schizosaccharomyces pombe. By obtaining 1-10 ng immunoprecipitated DNA, we generated genome-wide H3K9me2 or CENP-A/Cnp1 maps of the meiotic progeny from the genetic crossings of mhf2+(cen1_inactive) × mhf2+ (cen1_active) and mhf2? (cen1_inactive) × mhf2+ (cen1_active) in fission yeast. We find that the endogenous Centromere 1 in mhf2+ is converted to the inactivated Centromere 1 by mhf2? through genetic crossing. Overall design: 8 meiotic progeny (8 viable spores from 2 intact tetrads) from the genetic crossing of mhf2+ (cen1_inactive) × mhf2+ (cen1_active); 14 meiotic progeny (4 viable spores from 1 intact tetrad and 10 viable spores from 2 incomplete tetrads) from the genetic crossing of mhf2? (cen1_inactive) × mhf2+ (cen1_active) were examined by anti-H3K9me2 ChIP-seq. 4 meiotic progeny (4 viable spores from 1 intact tetrad) from the genetic crossing of mhf2? (cen1_inactive) × mhf2+ (cen1_active) were examined by anti-CENP-A/Cnp1 ChIP-seq.

本研究报道了粟酒裂殖酵母（Schizosaccharomyces pombe）中组蛋白修饰H3K9me2与组蛋白变体CENP-A/Cnp1的高通量谱分析。通过获取1~10 ng免疫沉淀DNA，我们对两组遗传杂交产生的减数分裂子代进行了全基因组H3K9me2或CENP-A/Cnp1图谱绘制：一组为mhf2+（cen1失活）× mhf2+（cen1活化），另一组为mhf2Δ（cen1失活）× mhf2+（cen1活化）。我们发现，通过遗传杂交，mhf2Δ可将mhf2+菌株中原生的着丝粒1转化为失活型着丝粒1。整体实验设计：针对遗传杂交组合mhf2+（cen1失活）× mhf2+（cen1活化），我们对其8株减数分裂子代（来自2个完整四分体的8个存活孢子）开展了抗H3K9me2染色质免疫沉淀测序（ChIP-seq）检测；针对遗传杂交组合mhf2Δ（cen1失活）× mhf2+（cen1活化），其中14株减数分裂子代（来自1个完整四分体的4个存活孢子与2个不完整四分体的10个存活孢子）接受了抗H3K9me2 ChIP-seq检测，另有4株减数分裂子代（来自1个完整四分体的4个存活孢子）接受了抗CENP-A/Cnp1 ChIP-seq检测。