Fine-tuning of the PAX-SIX-EYA-DACH network by multiple microRNAs controls embryo myogenesis.
Fine-tuning of the PAX-SIX-EYA-DACH network by multiple microRNAs controls embryo myogenesis.
复制标题
通过多种 microRNA 对 PAX-SIX-EYA-DACH 网络进行微调控制胚胎肌发生。
DOI：
10.1016/j.ydbio.2020.10.005
复制
发表时间：
2021
影响因子：
2.7
通讯作者：
Viaut C
中科院分区：
文献类型：
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作者：
Viaut C
MicroRNA（miRNAs）是一类短的非编码RNA，在转录后调节基因表达，在发育和疾病（包括肌肉疾病）中具有广泛的意义。测序技术和生物信息学的进步导致在脊椎动物和其他物种中鉴定出大量的miRNAs，然而，对于这些miRNAs中的许多具体作用尚未确定。LNA原位杂交揭示了富含体节的miRNAs的表达模式，在这里我们重点描述miR-128的功能。我们发现，在发育中的鸡体节中，miR-128的miR-128的miR-128介导的敲低（KD）对骨骼肌发生具有负面影响。计算分析将转录因子EYA 4确定为候选靶点，这与miR-128和EYA 4显示相似表达谱的观察结果一致。荧光素酶实验证实miR-128与EYA 43 ′UTR相互作用，体内实验也表明EYA 4受miR-128调控。EYA 4是PAX-SIX-EYA-DACH（PSED）转录因子网络的成员。因此，我们在SIX 1/4、EYA 1/2/3和DACH 1的3′UTR中鉴定了额外的候选miRNA结合位点。使用米兰达算法，我们发现了miR-128的位点，以及其他肌源性miRNA，miR-1a，miR-206和miR-133 a，其中一些被实验证实为功能性miRNA靶位点。我们的研究结果表明，miR-128通过直接靶向EYA 4并间接影响其他PSED成员（包括SIX 4和PAX 3）参与调节骨骼肌发生。因此，在miR-128敲低后观察到的对肌生成的抑制作用通过PAX 3的伴随敲低而被挽救。此外，我们表明，PSED网络的转录因子是由多种肌肉富集microRNA共同调节。
MicroRNAs (miRNAs), short non-coding RNAs, which act post-transcriptionally to regulate gene expression, are of widespread significance during development and disease, including muscle disease. Advances in sequencing technology and bioinformatics led to the identification of a large number of miRNAs in vertebrates and other species, however, for many of these miRNAs specific roles have not yet been determined. LNAin situhybridisation has revealed expression patterns of somite-enriched miRNAs, here we focus on characterising the functions of miR-128. We show that antagomiR-mediated knockdown (KD) of miR-128 in developing chick somites has a negative impact on skeletal myogenesis. Computational analysis identified the transcription factor EYA4 as a candidate target consistent with the observation that miR-128 andEYA4display similar expression profiles. Luciferase assays confirmed that miR-128 interacts with theEYA43′UTR.In vivoexperiments also suggest thatEYA4is regulated by miR-128. EYA4 is a member of the PAX-SIX-EYA-DACH (PSED) network of transcription factors. Therefore, we identified additional candidate miRNA binding sites in the 3′UTR ofSIX1/4,EYA1/2/3andDACH1. Using the miRanda algorithm, we found sites for miR-128, as well as for other myogenic miRNAs, miR-1a, miR-206 and miR-133a, some of these were experimentally confirmed as functional miRNA target sites. Our results reveal that miR-128 is involved in regulating skeletal myogenesis by directly targetingEYA4with indirect effects on other PSED members, includingSIX4andPAX3. Hence, the inhibitory effect on myogenesis observed after miR-128 knockdown was rescued by concomitant knockdown ofPAX3. Moreover, we show that the PSED network of transcription factors is co-regulated by multiple muscle-enriched microRNAs.