Differential expression of KRT83 regulated by the transcript factor CAP1 in Chinese Tan sheep
文献类型： 外文期刊
作者： Liu, Yufang 1 ; Kang, Xiaolong 2 ; Yang, Wanjie 1 ; Xie, Minggui 3 ; Zhang, Jibin 4 ; Fang, Meiying 1 ;
作者机构： 1.China Agr Univ, Dept Anim Genet & Breeding, MOA Lab Anim Genet & Breeding, Natl Engn Lab Anim Breeding,Coll Anim Sci & Techn, Beijing 100193, Peoples R China
2.Ningxia Univ, Coll Agr, Yinchuan 750021, Peoples R China
3.Jiangxi Acad Agr Sci, Inst Husb & Vet, Nanchang 330200, Jiangxi, Peoples R China
4.Iowa State Univ, Dept Anim Sci, Ames, IA 50010 USA
关键词： Chinese Tan sheep;KRT83;Electrophoretic mobility shift assay;CAP1
期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）
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收录情况： SCI
摘要： Keratin 83 (KRT83) is an important keratin protein in hair development. In this study, expression of KRT83 was compared among different tissues and between 1-month-old lambs and 48-month adult of Chinese Tan sheep, which showed different fleece phenotypes. The results showed that KRT83 was only expressed in skin, and KRT83 mRNA level in skin was significantly higher in Tan lambs than in adult sheep. To further understand the expression regulation of KRT83 by transcription factors in Tan sheep, amplified sequences coving different ranges of KRT83 promoter region were inserted into a pGL3-basic vector and then transfected into sheep primary fibroblast cells. Luciferase assay indicated that the sequence from -218 bp to -10 bp in the KRT83 promoter induced the highest transcription activity of the vector in the fibroblast cells. Transcription factor adenylate cyclase-associated protein 1 (CAP1) was predicted by online tools within this region. Electrophoretic mobility shift assay (EMSA) confirmed binding of the purified CAP1 protein to the target core region from -88 bp to -10 bp, because mutation in the target core sequence resulted in failure of CAP1 binding to the target region. Moreover, overexpression of CAP1 protein led to repression of the KRT83 promoter activity in sheep primary fibroblast cells, and expression of CAP1 was lower in lambs than in adult sheep. Therefore, we concluded that CAP1 is a key transcription factor involved in negative regulation of KRT83 expression in Tan sheep skin. Our study provides new insights into the transcriptional regulation of KRT83 and further hints of its critical role in curly hair phenotype in sheep. (C) 2017 Elsevier B.V. All rights reserved.
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