Department of Genetics, Albert Einstein College of Medicine, Bronx, NY, USA.
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY, USA.
Epigenetics Chromatin. 2023 Feb 18;16(1):8. doi: 10.1186/s13072-023-00481-y.
KDM5 family proteins are multi-domain regulators of transcription that when dysregulated contribute to cancer and intellectual disability. KDM5 proteins can regulate transcription through their histone demethylase activity in addition to demethylase-independent gene regulatory functions that remain less characterized. To expand our understanding of the mechanisms that contribute to KDM5-mediated transcription regulation, we used TurboID proximity labeling to identify KDM5-interacting proteins.
Using Drosophila melanogaster, we enriched for biotinylated proteins from KDM5-TurboID-expressing adult heads using a newly generated control for DNA-adjacent background in the form of dCas9:TurboID. Mass spectrometry analyses of biotinylated proteins identified both known and novel candidate KDM5 interactors, including members of the SWI/SNF and NURF chromatin remodeling complexes, the NSL complex, Mediator, and several insulator proteins.
Combined, our data shed new light on potential demethylase-independent activities of KDM5. In the context of KDM5 dysregulation, these interactions may play key roles in the alteration of evolutionarily conserved transcriptional programs implicated in human disorders.
KDM5 家族蛋白是转录的多结构域调节剂，当其失调时会导致癌症和智力障碍。KDM5 蛋白可以通过其组蛋白去甲基化酶活性以及去甲基化酶非依赖性的基因调控功能来调节转录，但其功能仍不太明确。为了扩大我们对 KDM5 介导的转录调控机制的理解，我们使用 TurboID 邻近标记来鉴定 KDM5 相互作用蛋白。
我们使用黑腹果蝇，通过使用新生成的 dCas9：TurboID 形式的 DNA 相邻背景对照，从 KDM5-TurboID 表达的成年头部中富集生物素化蛋白。生物素化蛋白的质谱分析鉴定了已知和新的候选 KDM5 相互作用蛋白，包括 SWI/SNF 和 NURF 染色质重塑复合物、NSL 复合物、Mediator 和几个绝缘子蛋白的成员。
综上所述，我们的数据为 KDM5 的潜在去甲基化酶非依赖性活性提供了新的线索。在 KDM5 失调的情况下，这些相互作用可能在改变与人类疾病相关的进化保守转录程序中发挥关键作用。
