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Differential expression of KRT83 regulated by the transcript factor CAP1 in Chinese Tan sheep
Gene (2017)
  • Yufang Liu, China Agricultural University
  • Xiaolong Kang, Ningxia University
  • Wanjie Yang, China Agricultural University
  • Minggui Xie, Jiangxi Academy of Agricultural Science
  • Jibin Zhang, Iowa State University
  • Meiying Fang, China Agricultrual University
Abstract
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.
Keywords
  • Chinese Tan sheep,
  • KRT83,
  • Electrophoretic mobility shift assay,
  • CAP1,
  • Fleece Development,
  • Gene expression regulation
Publication Date
Spring March 8, 2017
DOI
10.1016/j.gene.2017.03.007.
Citation Information
Yufang Liu, Xiaolong Kang, Wanjie Yang, Minggui Xie, et al.. "Differential expression of KRT83 regulated by the transcript factor CAP1 in Chinese Tan sheep" Gene Vol. 614 (2017) p. 15 - 20 ISSN: 0378-1119
Available at: http://works.bepress.com/jibin-zhang/17/