MC1R 5'-侧翼序列多态性对太行鸡羽色的影响。

Effect of polymorphisms in the 5'-flanking sequence of MC1R on feather color in Taihang chickens.

机构信息

College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056021, China.

Peking University Health Science Center, Peking University, Beijing, 100193, China.

出版信息

Poult Sci. 2022 Dec;101(12):102192. doi: 10.1016/j.psj.2022.102192. Epub 2022 Sep 29.

DOI:10.1016/j.psj.2022.102192

PMID:36283141

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9593195/

Abstract

MC1R plays an important role in the regulation of the formation, transfer, and deposition of melanin in animals and is important for determining coat color. Many studies have reported on single nucleotide polymorphisms (SNPs) in the coding sequence of MC1R. However, few studies have investigated the polymorphisms in the 5'-flanking sequence of MC1R. In this study, we sequenced 2000 bp of the 5'-flanking sequence of MC1R in 300 Taihang chickens with brown feathers (MTH) and 300 Taihang chickens with black feathers (HTH). The sequencing results showed that 4 SNPs (MC1R g.18838722 G > C, g.18838624 T > C, g.18838694 G > A, and g.18838624 C > T) were located in the 5'-flanking sequence of MC1R between the MTH and HTH groups. Association analysis showed that there was a significant correlation between the 4 SNPs and feather color in Taihang chickens. The correlation between MC1R g.18838624 T >C and feather color of Taihang chicken was 100%, of which the CC (E1) genotype is MTH and the TT (E2) genotype is HTH. Furthermore, there was a significant correlation between MC1R g.18838624 T > C and egg production at 302 d. E1 (184.14 ± 0.674) was significantly higher than that in E2 (181.75 ± 0.577) (P < 0.05). Luciferase reporter assays were used to detect the transcriptional activity of MC1R with different SNP genotypes. The results showed that the luciferase activity of E2 was significantly higher than that of E1 (P < 0.05). In addition, transcription factor-binding site predictions showed that E2 creates a new binding site for ZEB1. RT‒qPCR results revealed that the expression of MC1R in E2 was significantly lower than that in E1 (P < 0.05), and the expression of ZEB1 in E2 was significantly higher than that in E1 (P < 0.05). Overexpression and shRNA experiments demonstrated that ZEB1 regulates the expression of MC1R in DF1 cells. ZEB1 has a negative regulatory effect on the transcriptional activity of MC1R; it inhibits the expression of MC1R and affects the feather color of Taihang chickens. This study provides new insight into the molecular mechanism of feather color formation and the transcriptional regulation of MC1R in Taihang chickens.

摘要

MC1R 在动物中黑色素的形成、转移和沉积的调节中发挥着重要作用，对于决定毛色至关重要。许多研究报道了 MC1R 编码序列中的单核苷酸多态性（SNP）。然而，很少有研究调查 MC1R 5'-侧翼序列的多态性。在这项研究中，我们对 300 只棕色羽毛太行鸡（MTH）和 300 只黑色羽毛太行鸡（HTH）的 MC1R 的 5'-侧翼序列进行了 2000bp 的测序。测序结果表明，在 MTH 和 HTH 组之间的 MC1R 5'-侧翼序列中存在 4 个 SNP（MC1R g.18838722 G > C、g.18838624 T > C、g.18838694 G > A 和 g.18838624 C > T）。关联分析表明，这 4 个 SNP 与太行鸡的羽色存在显著相关性。MC1R g.18838624 T > C 与太行鸡羽色的相关性为 100%，其中 CC（E1）基因型为 MTH，TT（E2）基因型为 HTH。此外，MC1R g.18838624 T > C 与 302d 产蛋量显著相关。E1（184.14±0.674）显著高于 E2（181.75±0.577）（P < 0.05）。使用荧光素酶报告基因检测不同 SNP 基因型 MC1R 的转录活性。结果表明，E2 的荧光素酶活性显著高于 E1（P < 0.05）。此外，转录因子结合位点预测表明，E2 为 ZEB1 创造了一个新的结合位点。RT-qPCR 结果显示，E2 中 MC1R 的表达显著低于 E1（P < 0.05），E2 中 ZEB1 的表达显著高于 E1（P < 0.05）。过表达和 shRNA 实验表明，ZEB1 调节 DF1 细胞中 MC1R 的表达。ZEB1 对 MC1R 的转录活性具有负调控作用；它抑制 MC1R 的表达并影响太行鸡的羽色。本研究为太行鸡羽色形成的分子机制和 MC1R 的转录调控提供了新的见解。