Cell and Molecular Biology Lab, Veterinary Science Centre, University College Dublin, Dublin 4, Ireland.
Mol Biol Rep. 2012 Feb;39(2):919-28. doi: 10.1007/s11033-011-0817-z. Epub 2011 May 12.
Neuropeptide Y (NPY) is a potent orexigenic agent. The molecular mechanisms underlying the regulation of bovine NPY gene expression by its promoter region is currently unknown. The objectives of this research were to: (i) identify the SNPs in the promoter region of the bovine NPY gene, (ii) investigate the effects of these SNPs by measuring promoter transcriptional activities of different bovine NPY promoter haplotypes and; (iii) identify the minimal promoter region (MPR) required for basal activity of the NPY gene in vitro. Seventeen SNPs were identified in the promoter region. Of these, 14 affected putative transcription factors binding motifs including a TATA binding protein factor at -20, GC-Box factors SP1 at -170 and GATA binding motifs at -120 and -347. The SNPs were assigned to five major haplotypes (BtNPY_H1-5), of which BtNPY_H5 had maximum transcriptional activity. The region extending to -134 nt was identified as the MPR. This MPR was confirmed by the identification of a putative TATA box (-29 nt) and two SP1/GC binding sites (-94 and -118 nt), within this region. However, promoter expression was significantly enhanced when the construct contained the -614 to -1019 nt region. In conclusion, a number of SNPs characterised in the bovine NPY promoter especially those affecting the transcription factor binding sites, enhancer and repressor regions have the potential to affect NPY gene expression. Natural variation exists in the promoter region of the bovine NPY gene, which should be further explored for selection of energetic efficiency in cattle.
神经肽 Y（NPY）是一种有效的食欲刺激剂。目前尚不清楚其启动子区域调节牛 NPY 基因表达的分子机制。本研究的目的是：（i）鉴定牛 NPY 基因启动子区域中的 SNP；（ii）通过测量不同牛 NPY 启动子单倍型的启动子转录活性来研究这些 SNP 的影响；（iii）鉴定体外 NPY 基因基础活性所需的最小启动子区域（MPR）。在启动子区域鉴定出 17 个 SNP。其中，14 个 SNP 影响了假定的转录因子结合基序，包括 -20 处的 TATA 结合蛋白因子、-170 处的 SP1 GC 盒因子和 -120 和 -347 处的 GATA 结合基序。这些 SNP 被分为五个主要单倍型（BtNPY_H1-5），其中 BtNPY_H5 具有最大的转录活性。-134 nt 延伸区域被鉴定为 MPR。在该区域内，鉴定出了一个假定的 TATA 盒（-29 nt）和两个 SP1/GC 结合位点（-94 和 -118 nt），证实了这一 MPR。然而，当构建体包含-614 至-1019 nt 区域时，启动子表达显著增强。总之，牛 NPY 启动子中鉴定出的一些 SNP 特征，特别是那些影响转录因子结合位点、增强子和抑制剂区域的 SNP，可能会影响 NPY 基因的表达。牛 NPY 基因启动子区域存在自然变异，应进一步探索其在牛的能量效率选择中的应用。
