Cellular Memory Laboratory, RIKEN Cluster for Pioneering Research, Wako City, Saitama 351-0198, Japan.
Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA.
Nucleic Acids Res. 2024 Sep 23;52(17):10194-10219. doi: 10.1093/nar/gkae677.
The chromatin-remodeling enzyme helicase lymphoid-specific (HELLS) interacts with cell division cycle-associated 7 (CDCA7) on nucleosomes and is involved in the regulation of DNA methylation in higher organisms. Mutations in these genes cause immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome, which also results in DNA hypomethylation of satellite repeat regions. We investigated the functional domains of human CDCA7 in HELLS using several mutant CDCA7 proteins. The central region is critical for binding to HELLS, activation of ATPase, and nucleosome sliding activities of HELLS-CDCA7. The N-terminal region tends to inhibit ATPase activity. The C-terminal 4CXXC-type zinc finger domain contributes to CpG and hemimethylated CpG DNA preference for DNA-dependent HELLS-CDCA7 ATPase activity. Furthermore, CDCA7 showed a binding preference to DNA containing hemimethylated CpG, and replication-dependent pericentromeric heterochromatin foci formation of CDCA7 with HELLS was observed in mouse embryonic stem cells; however, all these phenotypes were lost in the case of an ICF syndrome mutant of CDCA7 mutated in the zinc finger domain. Thus, CDCA7 most likely plays a role in the recruitment of HELLS, activates its chromatin remodeling function, and efficiently induces DNA methylation, especially at hemimethylated replication sites.
染色质重塑酶解旋酶淋巴特异性 (HELLS) 在核小体上与细胞分裂周期相关蛋白 7 (CDCA7) 相互作用,参与高等生物中 DNA 甲基化的调控。这些基因的突变导致免疫缺陷、着丝粒不稳定和面部异常 (ICF) 综合征,这也导致卫星重复区域的 DNA 低甲基化。我们使用几种突变型 CDCA7 蛋白研究了人 CDCA7 在 HELLS 中的功能结构域。中央区域对于与 HELLS 的结合、ATP 酶的激活以及 HELLS-CDCA7 的核小体滑动活性至关重要。N 端区域倾向于抑制 ATP 酶活性。C 端 4CXXC 型锌指结构域有助于 CpG 和半甲基化 CpG DNA 对依赖 DNA 的 HELLS-CDCA7 ATP 酶活性的偏好。此外,CDCA7 显示出对含有半甲基化 CpG 的 DNA 的结合偏好,并且在小鼠胚胎干细胞中观察到 CDCA7 与 HELLS 之间复制依赖性着丝粒周围异染色质焦点的形成;然而,在锌指结构域发生 ICF 综合征突变的 CDCA7 情况下,所有这些表型都丢失了。因此,CDCA7 很可能在招募 HELLS 中发挥作用,激活其染色质重塑功能,并有效地诱导 DNA 甲基化,特别是在半甲基化的复制位点。