Novel regulatory sequence-82/-62 functions as a key element to drive the somite-specificity of zebrafish myf-5
Novel regulatory sequence-82/-62 functions as a key element to drive the somite-specificity of zebrafish myf-5
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DOI：
10.1002/dvdy.10357
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发表时间：
2003-09-01
影响因子：
2.5
通讯作者：
Tsai, HJ
中科院分区：
文献类型：
--
作者：
Chen, YH;Lee, HC;Tsai, HJ
MyF-5是一种控制肌肉分化的转录因子，在胚胎发育早期的体节中表达。然而，对Myf-5的基因调控知之甚少，需要对调控顺式元件进行详细的功能分析。在斑马鱼中，从-82到-62(-82/-62)的Myf-5上游序列与基本启动子融合，并转移到斑马鱼受精卵中。-82/-62盒驱动绿色荧光蛋白(GFP)报告基因在体节特异性表达。此外，在注射pCMV-5x(-82/-62)的28-HPF胚胎的体节中也能检测到GFP信号，其中含有5个拷贝的-82/-62盒插入巨细胞病毒启动子/增强子中。因此，在小鼠Myf-5中保守的-82/-62盒在斑马鱼胚胎的早期发育过程中具有驱动体节特异性表达和抑制非特异性表达的功能。对-82/-62盒的突变序列分析表明，-70/-62序列是控制Myf-5特异性的关键元件。推测的CCAAT样框位于-66/-62，不能指导体节特异性表达。在-70/-62探针和胚胎核提取液之间形成了DNA-蛋白质复合体。我们的结论是，-70/-62基序对于控制体节特异性表达是必不可少的，而CCAAT样框对于激活基因转录是必不可少的。发展动力学228.41-50,2003年。(C)2003年Wiley-Liss，Inc.
Myf-5, a transcription factor that controls muscle differentiation, is expressed in somites during early embryogenesis. However, gene regulation of myf-5 is poorly understood and detailed functional analysis of the regulatory cis-elements is needed. In zebrafish, the myf-5 upstream sequence from -82 to -62 (-82/-62) was fused with a basal promoter and transferred to fertilized zebrafish eggs. The -82/-62 cassette drove green fluorescent protein (GFP) reporter gene expression specifically in the somites. Moreover, GFP signals were detected exclusively in the somites of 28-hpf embryos derived from eggs injected with pCMV-5x(-82/-62), which contained five copies of the -82/-62 cassette inserted within cytomegalovirus promoter/enhancer. Thus, the -82/-62 cassette, conserved in mouse myf-5, functions to drive somite-specific expression and to repress nonspecific expression during the early development of zebrafish embryos. Mutated sequence analysis of -82/-62 cassette showed that the -70/-62 sequence was the key element for controlling myf-5 specificity. The putative CCAAT-like box, located at -66/-62, could not direct somite-specific expression. A DNA-protein complex was specifically formed between the -70/-62 probe and embryonic nuclear extracts. We conclude that the -70/-62 motif is essential for controlling somite-specific expression and the CCAAT-like box is essential for activating gene transcription. Developmental Dynamics 228.41-50, 2003. (C) 2003 Wiley-Liss, Inc.