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果蝇副肌球蛋白/小副肌球蛋白基因表达的调控。肌肉特异性转录的差异调控机制。

Control of Drosophila paramyosin/miniparamyosin gene expression. Differential regulatory mechanisms for muscle-specific transcription.

作者信息

Arredondo J J, Ferreres R M, Maroto M, Cripps R M, Marco R, Bernstein S I, Cervera M

机构信息

Departamento de Bioquímica & Instituto Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas, Facultad de Medicina, Universidad Autónoma de Madrid, Arzobispo Morcillo 4, 28029 Madrid, Spain.

出版信息

J Biol Chem. 2001 Mar 16;276(11):8278-87. doi: 10.1074/jbc.M009302200. Epub 2000 Nov 10.

Abstract

To define the transcriptional mechanisms contributing to stage- and tissue-specific expression of muscle genes, we performed transgenic analysis of Drosophila paramyosin gene regulation. This gene has two promoters, one for paramyosin and one for miniparamyosin, which are active in partially overlapping domains. Regions between -0.9 and -1.7 kilobases upstream of each initiation site contribute to the temporal and spatial expression patterns. By comparing the Drosophila melanogaster and Drosophila virilis promoters, conserved binding sites were found for known myogenic factors, including one MEF2 site and three E boxes. In contrast with previous data, our experiments with the paramyosin promoter indicate that the MEF2 site is essential but not sufficient for proper paramyosin gene transcription. Mutations in the three E boxes, on the other hand, do not produce any effect in embryonic/larval muscles. Thus MEF2 site- and E box-binding proteins can play different roles in the regulation of different muscle-specific genes. For the miniparamyosin promoters, several conserved sequences were shown to correspond to functionally important regions. Our data further show that the two promoters work independently. Even when both promoters are active in the same muscle fiber, the transcription driven by one of the promoters is not affected by transcription driven by the other.

摘要

为了确定促成肌肉基因阶段特异性和组织特异性表达的转录机制,我们对果蝇副肌球蛋白基因调控进行了转基因分析。该基因有两个启动子,一个用于副肌球蛋白,一个用于小副肌球蛋白,它们在部分重叠的结构域中起作用。每个起始位点上游-0.9至-1.7千碱基之间的区域促成了时间和空间表达模式。通过比较黑腹果蝇和 virilis 果蝇的启动子,发现了已知肌源性因子的保守结合位点,包括一个MEF2位点和三个E框。与先前的数据相反,我们用副肌球蛋白启动子进行的实验表明,MEF2位点对于副肌球蛋白基因的正确转录是必不可少的,但并不充分。另一方面,三个E框中的突变在胚胎/幼虫肌肉中没有产生任何影响。因此,MEF2位点结合蛋白和E框结合蛋白在不同肌肉特异性基因的调控中可以发挥不同的作用。对于小副肌球蛋白启动子,几个保守序列显示对应于功能重要区域。我们的数据进一步表明,这两个启动子独立工作。即使两个启动子在同一肌纤维中都处于活跃状态,其中一个启动子驱动的转录也不受另一个启动子驱动的转录的影响。

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