Base-Editing-Mediated R17H Substitution in Histone H3 Reveals Methylation-Dependent Regulation of Yap Signaling and Early Mouse Embryo Development

• Published in: Cell reports (Cambridge) 2019-01, Vol.26 (2), p.302-312.e4
• Main Authors: Yang, Guang, Zhou, Changyang, Wang, Ran, Huang, Shisheng, Wei, Yu, Yang, Xianfa, Liu, Yajing, Li, Jianan, Lu, Zongyang, Ying, Wenqin, Li, Xiajun, Jing, Naihe, Huang, Xingxu, Yang, Hui, Qiao, Yunbo
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - metabolism; Animals; base editing; CARM1; Catalytic Domain; Cell Cycle; Cell Cycle Proteins - metabolism; Cell Line, Tumor; embryo development; Embryonic Development; Gene Expression Regulation, Developmental; H3R17; Histone Code; histone methylation; Histones - chemistry; Histones - genetics; Histones - metabolism; Mice; Mice, Inbred C57BL; Mice, Inbred ICR; Mutation, Missense; Protein-Arginine N-Methyltransferases - chemistry; Protein-Arginine N-Methyltransferases - genetics; Protein-Arginine N-Methyltransferases - metabolism; Signal Transduction; Transcriptome; Yap1 signaling
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2018.12.046

The coactivator-associated arginine methyltransferase CARM1 catalyzes the methylation of histone H3 arginine 17/26 (H3R17/26me) and non-histone proteins at arginine residues to regulate gene transactivation through profiling or Carm1 overexpression assays. However, the direct relationship between H3R17/26me and its causal role in mouse embryo development remains largely unclear. Here, we use rAPOBEC1-XTEN-Cas9n-UGI (BE3) to efficiently introduce a point mutation (R17H) at multiple Hist1/2H3 loci and a premature-stop codon into the catalytic domain of CARM1 in mouse embryos, resulting in remarkable downregulation of H3R17me levels and developmental defects in pre-implantation and fetal embryos. Transcriptomic analysis reveals that Yap1 and cell cycle signaling pathways are dysregulated in Carm1 truncation and H3R17H substitution embryos, and Yap1 overexpression could rescue the base-editing-elicited defects. Our data establish the direct regulatory relationship between CARM1-mediated H3R17me and early mouse embryo development and demonstrate that Yap1 acts downstream of CARM1-mediated H3R17me to regulate the mouse embryo development. [Display omitted] •Base editing introduces substitutions at multiple histone H3 loci•Both H3R17H substitution and Carm1 truncation disrupt early embryo development•H3R17H substitution and Carm1 truncation lead to similar transcriptomic alterations•H3R17me regulates embryonic development through Yap1 and cell cycle signaling pathways Yang et al. apply CRISPR-Cas9-based base editing tools to precisely introduce substitutions at multiple histone H3 loci to disrupt methylation at histone H3 arginine 17 (H3R17me), and they find that CARM1-mediated H3R17me regulates embryo development through Yap1 and cell cycle signaling pathways.