cis-Regulatory elements conserved in the proximal promoter region of an ascidian embryonic muscle myosin heavy-chain gene

cis-Regulatory elements conserved in the proximal promoter region of an ascidian embryonic muscle myosin heavy-chain gene
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海鞘胚胎肌肌球蛋白重链基因近端启动子区保守的顺式调控元件

DOI:
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发表时间:
1996
期刊:
Development, Genes and Evolution
影响因子:
--
通讯作者:
Noriyuki Satoh
Noriyuki Satoh
中科院分区:
--
文献类型:
--
作者:
I. Araki;Noriyuki Satoh

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摘要海鞘胚胎的B-线肌细胞是根据未受精卵肌质中预先定位的决定子自主指定的。 肌肉特异性肌动蛋白和肌球蛋白重链基因的表达早在32细胞期就开始于B系假定肌细胞。为了探索这种分化的内在遗传程序,我们分析了罗氏盐辛西娅肌肉肌球蛋白重链基因(HrMHC 1)的顺式调控元件。HrMHC 1与其他无脊椎动物和脊椎动物的氨基酸序列的比较表明,HrMHC 1类似于脊椎动物骨骼肌和心肌的肌球蛋白重链。将HrMHC 1基因5′-端上游132 bp与细菌lacZ报告基因融合,构建了HrMHC 1融合基因。 将融合基因显微注射入受精卵后,报告基因最终仅在尾芽胚胎的肌细胞中表达。据报道,海鞘胚胎肌肉肌动蛋白基因(HrMA 4)的转录起始位点的序列5′的103 bp包含足够用于肌肉特异性表达的信息(Hikoietet al.  1994年)。比较HrMHC 1基因5′端132 bp序列和HrMA 4基因5′端103 bp序列,发现两个基因有几个共同的基序(E-box、加塔box和Boxes A、B、T1和T2)。  插入这些基序的点突变表明,盒T1/T2(TTTTTTCTTTCA)是关键的HrMHC 1基因的启动子活性。
Abstract The B-line muscle cells of the ascidian embryo are specified autonomously depending on determinants prelocalized in the myoplasm of unfertilized eggs. Expression of muscle-specific actin and myosin heavy-chain genes commences in the B-line presumptive muscle cells as early as the 32-cell stage. To explore the intrinsic genetic program for this differentiation, we analysed cis-regulatory elements of the Halocynthia roretzi muscle myosin heavy-chain gene (HrMHC1). Comparison of the entire amino acid sequence of HrMHC1 with those of other invertebrates and vertebrates indicated that HrMHC1 resembles myosin heavy-chain of vertebrate skeletal and cardiac muscles. A fusion gene was constructed consisting of 132 bp upstream the 5′-end of HrMHC1 gene fused to a bacterial lacZ reporter. When the fusion gene was microinjected into fertilized eggs, the reporter gene was eventually expressed only in muscle cells of tailbud embryos. It has been reported that 103 bp of sequence 5′ of the transcription start site of the ascidian embryonic muscle actin gene (HrMA4) contains information sufficient for muscle-specific expression (Hikosaka et al. 1994). Comparison of the 132 bp of sequence 5′ of the HrMHC1 gene with the 103 bp of sequence 5′ of the HrMA4 gene revealed several common motifs shared by the two genes (E-box, GATA box and Boxes A, B, T1 and T2). Point mutations inserted into these motifs suggested that the Box T1/T2 (TTTTTTCTTTCA) is critical for the promoter activity of the HrMHC1 gene.
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