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非洲爪蟾发育过程中中间丝蛋白的表达。II. 结蛋白的鉴定与分子特征分析。

Expression of intermediate filament proteins during development of Xenopus laevis. II. Identification and molecular characterization of desmin.

作者信息

Herrmann H, Fouquet B, Franke W W

机构信息

Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.

出版信息

Development. 1989 Feb;105(2):299-307. doi: 10.1242/dev.105.2.299.

Abstract

During embryogenesis of avian and mammalian species the formation of intermediate filaments (IFs) containing desmin is characteristic for myogenesis. In view of important differences of patterns of IF protein expression in embryogenic pathways of amphibia on the one hand and birds and mammals on the other, we have decided to study the expression of desmin during early embryogenesis of Xenopus laevis by cDNA hybridization and antibody reactions. Here we describe the isolation of a cDNA clone encoding Xenopus desmin and the deduced amino acid sequence (458 residues; Mr 52,800) which displays a very high degree of conservation during vertebrate evolution from Xenopus to chicken and hamster, with a similar degree of sequence divergence between all three species compared. In addition, we have noted, by both cDNA-hybrid-selection-translation and immunoblotting of cytoskeletal proteins a second desmin-related polypeptide of Mr approximately 49,000. RNA (Northern) blot analyses show the occurrence of three different desmin mRNAs (1.9, 2.6 and 3.0 kb) which seem to represent different polyadenylation sites, displaying quantitative differences in different kinds of muscle tissues. During embryogenesis, desmin mRNA has first been detected in stage-14 embryos and then increases drastically to high levels at stage 18 and thereafter. Immunofluorescence microscopy using desmin-specific antibodies shows that this synthesis of desmin is restricted to somite tissue. The embryonic time course of synthesis of desmin and desmin mRNA is discussed in relation to those of other muscle proteins.

摘要

在鸟类和哺乳动物的胚胎发生过程中,含有结蛋白的中间丝(IFs)的形成是肌发生的特征。鉴于两栖动物与鸟类和哺乳动物在胚胎发生途径中IF蛋白表达模式存在重要差异,我们决定通过cDNA杂交和抗体反应研究非洲爪蟾早期胚胎发生过程中结蛋白的表达。在此,我们描述了一个编码非洲爪蟾结蛋白的cDNA克隆的分离及其推导的氨基酸序列(458个残基;Mr 52,800),该序列在从非洲爪蟾到鸡和仓鼠的脊椎动物进化过程中显示出高度的保守性,与所有这三个物种之间的序列差异程度相似。此外,我们通过cDNA杂交选择翻译和细胞骨架蛋白的免疫印迹法都注意到了一种Mr约为49,000的第二种结蛋白相关多肽。RNA(Northern)印迹分析显示存在三种不同的结蛋白mRNA(1.9、2.6和3.0 kb),它们似乎代表不同的多聚腺苷酸化位点,在不同类型的肌肉组织中表现出定量差异。在胚胎发生过程中,结蛋白mRNA首先在14期胚胎中被检测到,然后在18期及之后急剧增加到高水平。使用结蛋白特异性抗体的免疫荧光显微镜检查表明,结蛋白的这种合成仅限于体节组织。本文结合其他肌肉蛋白的合成过程讨论了结蛋白和结蛋白mRNA合成的胚胎时间进程。

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