Cell-autonomous and redundant roles of Hey1 and HeyL in muscle stem cells: HeyL requires Hes1 to bind diverse DNA sites
Cell-autonomous and redundant roles of Hey1 and HeyL in muscle stem cells: HeyL requires Hes1 to bind diverse DNA sites
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DOI：
10.1242/dev.163618
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发表时间：
2019-02
期刊：
影响因子：
4.6
通讯作者：
Yu-taro Noguchi;Miki Nakamura;Nobumasa Hino;Jumpei Nogami;Sayaka Tsuji;Takahiko Sato;Lidan Zhang;K. Tsujikawa;Toru Tanaka;K. Izawa;Yoshiaki Okada;T. Doi;H. Kokubo;Akihito Harada;A. Uezumi;M. Gessler;Y. Ohkawa;S. Fukada
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文献类型：
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作者：
Yu-taro Noguchi;Miki Nakamura;Nobumasa Hino;Jumpei Nogami;Sayaka Tsuji;Takahiko Sato;Lidan Zhang;K. Tsujikawa;Toru Tanaka;K. Izawa;Yoshiaki Okada;T. Doi;H. Kokubo;Akihito Harada;A. Uezumi;M. Gessler;Y. Ohkawa;S. Fukada
摘要肌干（卫星）细胞（MuSCs）的未分化状态由经典Notch通路维持。虽然三个bHLH转录因子，Hey 1，HeyL和Hes 1，被认为是Notch途径发挥抗肌生成作用的潜在效应子，但HeyL和Hes 1均不抑制肌生成细胞系的肌生成分化。此外，这些因素是否冗余或合作工作是未知的。在这里，我们使用条件和遗传无效小鼠显示了Hey 1和HeyL在MuSC中的细胞自主功能。对培养的MuSC的分析揭示了HeyL和Hes 1两者的抗肌生成活性。我们发现HeyL在活细胞中与Hes 1形成异二聚体复合物。此外，我们的ChIP-seq实验表明，与单独的HeyL相比，HeyL-Hes 1异二聚体以高亲和力结合染色质中的特定位点，包括Hey 1的结合位点。最后，肌细胞生成素启动子活性的分析表明，HeyL和Hes 1协同作用，抑制肌细胞分化。总的来说，这些结果表明HeyL和Hey 1在MuSC中具有冗余功能，并且HeyL需要Hes 1来进行有效的DNA结合和生物活性。总结：对条件性和遗传无效小鼠的分析表明，HeyL和Hey 1在MuSC中具有冗余功能，并且HeyL需要Hes 1才能有效地结合DNA和发挥生物活性。
ABSTRACT The undifferentiated state of muscle stem (satellite) cells (MuSCs) is maintained by the canonical Notch pathway. Although three bHLH transcriptional factors, Hey1, HeyL and Hes1, are considered to be potential effectors of the Notch pathway exerting anti-myogenic effects, neither HeyL nor Hes1 inhibits myogenic differentiation of myogenic cell lines. Furthermore, whether these factors work redundantly or cooperatively is unknown. Here, we showed cell-autonomous functions of Hey1 and HeyL in MuSCs using conditional and genetic null mice. Analysis of cultured MuSCs revealed anti-myogenic activity of both HeyL and Hes1. We found that HeyL forms heterodimeric complexes with Hes1 in living cells. Moreover, our ChIP-seq experiments demonstrated that, compared with HeyL alone, the HeyL-Hes1 heterodimer binds with high affinity to specific sites in the chromatin, including the binding sites of Hey1. Finally, analyses of myogenin promoter activity showed that HeyL and Hes1 act synergistically to suppress myogenic differentiation. Collectively, these results suggest that HeyL and Hey1 function redundantly in MuSCs, and that HeyL requires Hes1 for effective DNA binding and biological activity. Summary: Analyses of conditional and genetic null mice reveal that HeyL and Hey1 function redundantly in MuSCs, and that HeyL requires Hes1 for effective DNA binding and biological activity.