Department of Ecology and Evolutionary Biology, University of California, Irvine, California, United States of America.
Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, United States of America.
PLoS Genet. 2023 May 5;19(5):e1010439. doi: 10.1371/journal.pgen.1010439. eCollection 2023 May.
We use ATAC-seq to examine chromatin accessibility for four different tissues in Drosophila melanogaster: adult female brain, ovaries, and both wing and eye-antennal imaginal discs from males. Each tissue is assayed in eight different inbred strain genetic backgrounds, seven associated with a reference quality genome assembly. We develop a method for the quantile normalization of ATAC-seq fragments and test for differences in coverage among genotypes, tissues, and their interaction at 44099 peaks throughout the euchromatic genome. For the strains with reference quality genome assemblies, we correct ATAC-seq profiles for read mis-mapping due to nearby polymorphic structural variants (SVs). Comparing coverage among genotypes without accounting for SVs results in a highly elevated rate (55%) of identifying false positive differences in chromatin state between genotypes. After SV correction, we identify 1050, 30383, and 4508 regions whose peak heights are polymorphic among genotypes, among tissues, or exhibit genotype-by-tissue interactions, respectively. Finally, we identify 3988 candidate causative variants that explain at least 80% of the variance in chromatin state at nearby ATAC-seq peaks.
我们使用 ATAC-seq 技术检测了黑腹果蝇(Drosophila melanogaster)的四个不同组织的染色质可及性:成年雌性大脑、卵巢,以及雄性的翅膀和眼触角体节盘。每种组织在 8 种不同的近交系遗传背景下进行检测,其中 7 种与参考质量基因组组装有关。我们开发了一种用于 ATAC-seq 片段定量归一化的方法,并在整个常染色质基因组的 44099 个峰中测试了基因型、组织以及它们之间相互作用的覆盖差异。对于具有参考质量基因组组装的品系,我们纠正了由于附近多态性结构变异(SVs)而导致的读映射错误的 ATAC-seq 图谱。如果不考虑 SVs 来比较基因型之间的覆盖差异,会导致假阳性染色质状态差异的识别率(55%)非常高。在 SV 校正后,我们分别鉴定出了 1050、30383 和 4508 个峰,其峰高在基因型之间、组织之间或表现出基因型-组织相互作用中具有多态性。最后,我们鉴定出了 3988 个候选因果变异,它们至少解释了附近 ATAC-seq 峰处染色质状态方差的 80%。