Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.
Dev Cell. 2009 Dec;17(6):836-48. doi: 10.1016/j.devcel.2009.10.011.
We created a whole-mount in situ hybridization (WISH) database, termed EMBRYS, containing expression data of 1520 transcription factors and cofactors expressed in E9.5, E10.5, and E11.5 mouse embryos--a highly dynamic stage of skeletal myogenesis. This approach implicated 43 genes in regulation of embryonic myogenesis, including a transcriptional repressor, the zinc-finger protein RP58 (also known as Zfp238). Knockout and knockdown approaches confirmed an essential role for RP58 in skeletal myogenesis. Cell-based high-throughput transfection screening revealed that RP58 is a direct MyoD target. Microarray analysis identified two inhibitors of skeletal myogenesis, Id2 and Id3, as targets for RP58-mediated repression. Consistently, MyoD-dependent activation of the myogenic program is impaired in RP58 null fibroblasts and downregulation of Id2 and Id3 rescues MyoD's ability to promote myogenesis in these cells. Our combined, multi-system approach reveals a MyoD-activated regulatory loop relying on RP58-mediated repression of muscle regulatory factor (MRF) inhibitors.
我们创建了一个全胚胎原位杂交（WISH）数据库，称为 EMBRYS，其中包含了在 E9.5、E10.5 和 E11.5 小鼠胚胎中表达的 1520 个转录因子和辅助因子的表达数据，这是骨骼肌发生的一个高度动态的阶段。这种方法暗示了 43 个基因在胚胎肌发生中的调节作用，包括一个转录抑制因子，锌指蛋白 RP58（也称为 Zfp238）。敲除和敲低方法证实了 RP58 在骨骼肌发生中的重要作用。基于细胞的高通量转染筛选揭示了 RP58 是 MyoD 的直接靶标。微阵列分析确定了两个抑制骨骼肌发生的抑制剂，Id2 和 Id3，作为 RP58 介导的抑制的靶标。一致地，RP58 缺失成纤维细胞中 MyoD 依赖的肌发生程序的激活受损，下调 Id2 和 Id3 挽救了 MyoD 在这些细胞中促进肌发生的能力。我们的综合多系统方法揭示了一个 MyoD 激活的调节环，依赖于 RP58 介导的肌肉调节因子（MRF）抑制剂的抑制。
