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非洲爪蟾新的表皮角蛋白基因:无尾类皮肤变态过程中其表达的激素和区域调控

New epidermal keratin genes from Xenopus laevis: hormonal and regional regulation of their expression during anuran skin metamorphosis.

作者信息

Watanabe Y, Kobayashi H, Suzuki K, Kotani K, Yoshizato K

机构信息

Yoshizato MorphoMatrix Project, ERATO, JST and Developmental Biology Laboratory, Department of Biological Science, Graduate School of Science, Hiroshima University, Higashihiroshima, 739-8526, Hiroshima, Japan.

出版信息

Biochim Biophys Acta. 2001 Feb 16;1517(3):339-50. doi: 10.1016/s0167-4781(00)00281-5.

Abstract

Xenopus larval keratin (XLK) was isolated by gel electrophoresis of proteins of tadpole skin. Screening of an expression cDNA library of tail tissues by specific polyclonal antibodies against XLK produced XLK cDNA (xlk). Its complete nucleotide and predicted amino acid sequences revealed that XLK was a new member of type II keratin. Screening of a cDNA library of adult Xenopus skin using an oligonucleotide probe which had been designed from well-conserved N-terminal amino acid sequences of the rod domain of type I keratin produced two cDNAs, xak-a and xak-b, which were found to be new members of type I keratin gene. Northern blot analysis showed that xlk was expressed exclusively in the larval skin whereas xak-a and xak-b were expressed exclusively in the adult skin. Their expression level was regulated in a region- and metamorphic stage- dependent manner during larval skin development. mRNA in situ hybridization experiments identified the cells that expressed xlk, and xak-a and xak-b as larva- specific epidermal cells (skein cells and basal cells), and adult suprabasal epidermal cells, respectively. These three genes were found to be late responsive to thyroid hormone. Phylogenetic relationships of these keratins with known ones are discussed.

摘要

通过对蝌蚪皮肤蛋白质进行凝胶电泳分离出非洲爪蟾幼体角蛋白(XLK)。用针对XLK的特异性多克隆抗体筛选尾组织的表达cDNA文库,得到了XLK cDNA(xlk)。其完整的核苷酸序列和预测的氨基酸序列表明,XLK是II型角蛋白的一个新成员。使用根据I型角蛋白杆状结构域保守的N端氨基酸序列设计的寡核苷酸探针筛选成年非洲爪蟾皮肤的cDNA文库,得到了两个cDNA,即xak-a和xak-b,它们被发现是I型角蛋白基因的新成员。Northern印迹分析表明,xlk仅在幼体皮肤中表达,而xak-a和xak-b仅在成年皮肤中表达。在幼体皮肤发育过程中,它们的表达水平以区域和变态阶段依赖的方式受到调节。mRNA原位杂交实验确定了表达xlk、xak-a和xak-b的细胞分别为幼体特异性表皮细胞(细丝细胞和基底细胞)和成体基底上层表皮细胞。发现这三个基因对甲状腺激素反应较晚。讨论了这些角蛋白与已知角蛋白的系统发育关系。

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