NRF1 和 ZSCAN10 与 SIX1 基因的启动子区域结合并影响秦川牛的体测量。,Scientific Reports

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NRF1 和 ZSCAN10 与 SIX1 基因的启动子区域结合并影响秦川牛的体测量。
Scientific Reports ( IF 3.8 ) Pub Date : 2017-Aug-11 , DOI: 10.1038/s41598-017-08384-1
Da-Wei Wei _{1,

2} , Lin-Sheng Gui _{1,

2} , Sayed Haidar Abbas Raza ₁ , Song Zhang _{1,

2} , Rajwali Khan ₁ , Li Wang _{1,

2} , Hong-Fang Guo _{1,

2} , Lin-Sen Zan _{1,

3,

4}

Affiliation

SIX1 同源框基因属于六个同源域家族，被广泛认为在介导骨骼肌发育中起主要作用。在本研究中，我们确定牛 SIX1 基因在胸最长肌和生理不成熟的个体中高度表达。对 428 头秦川牛进行 DNA 测序，在 SIX1 基因的启动子区域鉴定出 9 个单核苷酸多态性 (SNP)。使用一系列 5' 缺失启动子质粒荧光素酶报告基因分析和 5'-快速扩增 cDNA 末端分析 (RACE)，发现这些 SNP 中的两个位于相对于转录的近端最小启动子区域 -216/-28起始点 (TSS)。相关分析显示组合单倍型 H ₁ -H ₂(-GG-GA-)在体测性状(BMTs)中显着大于其他，这与显示Hap2在秦川牛成肌细胞中的转录活性高于其他的结果一致。此外，电泳迁移率变动分析 (EMSA) 和染色质免疫沉淀分析 (ChIP) 表明 NRF1 和 ZSCAN10 结合发生在双倍型 H ₁ -H ₂的启动子区域以调节 SIX1 转录活性。该信息可能对牛育种中的分子标记辅助选择 (MAS) 有用。

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NRF1 and ZSCAN10 bind to the promoter region of the SIX1 gene and their effects body measurements in Qinchuan cattle.

The SIX1 homeobox gene belongs to the six homeodomain family and is widely thought to play a principal role in mediating of skeletal muscle development. In the present study, we determined that the bovine SIX1 gene was highly expressed in the longissimus thoracis and physiologically immature individuals. DNA sequencing of 428 individual Qinchuan cattle identified nine single nucleotide polymorphisms (SNPs) in the promoter region of the SIX1 gene. Using a series of 5' deletion promoter plasmid luciferase reporter assays and 5'-rapid amplification of cDNA end analysis (RACE), two of these SNPs were found to be located in the proximal minimal promoter region -216/-28 relative to the transcriptional start site (TSS). Correlation analysis showed the combined haplotypes H₁-H₂ (-GG-GA-) was significantly greater in the body measurement traits (BMTs) than the others, which was consistent with the results showing that the transcriptional activity of Hap2 was higher than the others in Qinchuan cattle myoblast cells. Furthermore, the electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation assay (ChIP) demonstrated that NRF1 and ZSCAN10 binding occurred in the promoter region of diplotypes H₁-H₂ to regulate SIX1 transcriptional activity. This information may be useful for molecular marker-assisted selection (MAS) in cattle breeding.

更新日期：2017-08-11

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