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牛 SIX1 基因启动子区的特征：MyoD、PAX7、CREB 和 MyoG 的作用。

Characterization of the promoter region of the bovine SIX1 gene: Roles of MyoD, PAX7, CREB and MyoG.

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, People's Republic of China.

Modern Cattle Biotechnology and Application of National-Local Engineering Research Center, Yangling, 712100, Shaanxi, People's Republic of China.

出版信息

Sci Rep. 2017 Oct 3;7(1):12599. doi: 10.1038/s41598-017-12787-5.

DOI:10.1038/s41598-017-12787-5
PMID:28974698
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5626756/
Abstract

The SIX1 gene belongs to the family of six homeodomain transcription factors (TFs), that regulates the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway and mediate skeletal muscle growth and regeneration. Previous studies have demonstrated that SIX1 is positively correlated with body measurement traits (BMTs). However, the transcriptional regulation of SIX1 remains unclear. In the present study, we determined that bovine SIX1 was highly expressed in the longissimus thoracis. To elucidate the molecular mechanisms involved in bovine SIX1 regulation, 2-kb of the 5' regulatory region were obtained. Sequence analysis identified neither a consensus TATA box nor a CCAAT box in the 5' flanking region of bovine SIX1. However, a CpG island was predicted in the region -235 to +658 relative to the transcriptional start site (TSS). An electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay in combination with serial deletion constructs of the 5' flanking region, site-directed mutation and siRNA interference demonstrated that MyoD, PAX7 and CREB binding occur in region -689/-40 and play important roles in bovine SIX1 transcription. In addition, MyoG drives SIX1 transcription indirectly via the MEF3 motif. Taken together these interactions suggest a key functional role for SIX1 in mediating skeletal muscle growth in cattle.

摘要

SIX1 基因属于六类同源结构域转录因子（TFs）家族，可调节细胞外信号调节激酶 1/2（ERK1/2）通路，并介导骨骼肌生长和再生。先前的研究表明，SIX1 与体尺性状（BMTs）呈正相关。然而，SIX1 的转录调控尚不清楚。在本研究中，我们确定牛 SIX1 在胸最长肌中高度表达。为了阐明牛 SIX1 调节的分子机制，我们获得了 5'调控区的 2kb 序列。序列分析未在牛 SIX1 的 5'侧翼区识别出一致的 TATA 盒或 CCAAT 盒。然而，在相对于转录起始位点（TSS）的-235 到+658 区域预测到一个 CpG 岛。电泳迁移率变动分析（EMSA）和染色质免疫沉淀（ChIP）分析结合 5'侧翼区的串联缺失构建体、定点突变和 siRNA 干扰表明，MyoD、PAX7 和 CREB 结合发生在-689/-40 区域，并在牛 SIX1 转录中发挥重要作用。此外，MyoG 通过 MEF3 基序间接驱动 SIX1 转录。这些相互作用表明 SIX1 在介导牛的骨骼肌生长中起着关键的功能作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/2563ccd8b0f9/41598_2017_12787_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/6ea11d2df0a2/41598_2017_12787_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/3d76a732b91b/41598_2017_12787_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/4fd4abac3612/41598_2017_12787_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/273085cfb774/41598_2017_12787_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/ddb0d3026b78/41598_2017_12787_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/2aeed1782135/41598_2017_12787_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/2563ccd8b0f9/41598_2017_12787_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/6ea11d2df0a2/41598_2017_12787_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/3d76a732b91b/41598_2017_12787_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/4fd4abac3612/41598_2017_12787_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/273085cfb774/41598_2017_12787_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/ddb0d3026b78/41598_2017_12787_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/2aeed1782135/41598_2017_12787_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f2b/5626756/2563ccd8b0f9/41598_2017_12787_Fig7_HTML.jpg

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