Re-analysis of publicly available methylomes using signal detection yields new information.

Re-analysis of publicly available methylomes using signal detection yields new information.
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DOI:
10.1038/s41598-023-30422-4
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发表时间:
2023-02-27
期刊:
影响因子:
4.6
通讯作者:
Mackenzie, Sally
Mackenzie, Sally
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Hafner, Alenka;Mackenzie, Sally

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胞嘧啶甲基化是一种表观遗传标记,参与植物基因表达和染色质稳定性的调节。全基因组测序技术的进步使研究不同条件下的甲基组动态成为可能。然而,分析亚硫酸氢盐序列数据的计算方法并不统一。争论仍然存在于差异甲基化位置与所研究的处理和排除噪声的相关性中,这是这些随机数据集固有的。流行的方法应用费舍尔精确检验、逻辑回归或贝塔回归,然后对甲基化水平的差异进行任意的截断。另一种不同的策略,即甲基IT管道,利用信号检测来根据甲基化发散的广义伽马概率分布来确定截止日期。对拟南芥两项表观遗传学研究中公开的BS-SEQ数据进行重新分析,并应用甲基IT技术,发现了以前未报道的其他结果。甲基组重组对磷酸盐饥饿的反应被证实是组织特异性的,除了最初研究中没有涉及的硫酸盐代谢基因外,还包括磷酸盐同化基因。在种子萌发过程中,植物经历了主要的甲基组重新编程,而甲基IT的使用使我们能够识别特定阶段的基因网络。我们从这些比较研究中推测,稳健的甲基组实验必须考虑到数据的随机性,才能实现有意义的功能分析。
Cytosine methylation is an epigenetic mark that participates in regulation of gene expression and chromatin stability in plants. Advancements in whole genome sequencing technologies have enabled investigation of methylome dynamics under different conditions. However, the computational methods for analyzing bisulfite sequence data have not been unified. Contention remains in the correlation of differentially methylated positions with the investigated treatment and exclusion of noise, inherent to these stochastic datasets. The prevalent approaches apply Fisher’s exact test, logistic, or beta regression, followed by an arbitrary cut-off for differences in methylation levels. A different strategy, the MethylIT pipeline, utilizes signal detection to determine cut-off based on a fitted generalized gamma probability distribution of methylation divergence. Re-analysis of publicly available BS-seq data from two epigenetic studies in Arabidopsis and applying MethylIT revealed additional, previously unreported results. Methylome repatterning in response to phosphate starvation was confirmed to be tissue-specific and included phosphate assimilation genes in addition to sulfate metabolism genes not implicated in the original study. During seed germination plants undergo major methylome reprogramming and use of MethylIT allowed us to identify stage-specific gene networks. We surmise from these comparative studies that robust methylome experiments must account for data stochasticity to achieve meaningful functional analyses.
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在多因素全基因组亚硫酸盐测序实验中,使用β-二项式回归进行高精度差甲基化分析。
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