Department of Animal Genetic, Breeding and Reproduction, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
Department of Animal Genetic and Breeding, College of Animal Production, University of Bahri, 1660, Khartoum North, Sudan.
Cell Stress Chaperones. 2018 Jul;23(4):639-651. doi: 10.1007/s12192-017-0873-y. Epub 2018 Jan 20.
The heat shock protein 90 (Hsp90) is a copious and ubiquitous molecular chaperone which plays an essential role in many cellular biological processes. The objective of this study was to identify single nucleotide polymorphisms (SNPs) in the Hsp90AA1 gene and to determine their association with heat stress traits in Chinese Holstein cattle breed. Direct sequencing was used to identify new SNPs. Luciferase reporter assay methods were used to assess g.- 87G > C and g.4172A > G loci in the promoter activity and 3'-UTR, respectively. Quantitative real-time PCR was utilized to quantify the gene expression profile. Five SNPs were identified in 130 multiparous lactating cows: one SNP in the promoter, three SNPs in the coding region, and one in 3'-UTR were novel and reported for the first time in this study. As a result of promoter assay using dual luciferase assay system, the genotype CC showed the highest transcription activity region (13.67 ± 0.578) compared to the wild-type GG (3.24 ± 0.103). On the other hand, the result revealed that one of the selected microRNAs (dme-miR-2279-5p) was found to interact with the Hsp90AA1 3'-UTR sequence and to suppress the reporter activity markedly in the presence of the allele G (2.480 ± 0.136). The expression of Hsp90AA1 in cow bearing mutant allele C was higher (4.18 ± 0.928) than cows bearing wild-type allele G (1.008 ± 0.0.129) in stress season. In summary, there was an association between genetic variations in the Hsp90AA1 and thermoresistance. This association could be used as a marker in genetic selection for heat tolerance in Chinese Holstein cattle breeds.
热休克蛋白 90（Hsp90）是一种丰富且普遍存在的分子伴侣，在许多细胞生物学过程中发挥着重要作用。本研究的目的是鉴定 Hsp90AA1 基因中的单核苷酸多态性（SNP），并确定它们与中国荷斯坦奶牛耐热性状的关系。直接测序用于鉴定新的 SNP。荧光素酶报告基因检测方法分别用于评估启动子活性和 3'-UTR 中的 g.-87G>C 和 g.4172A>G 位点。实时定量 PCR 用于量化基因表达谱。在 130 头经产泌乳奶牛中鉴定出 5 个 SNP：启动子中 1 个 SNP、编码区中 3 个 SNP 和 3'-UTR 中 1 个 SNP 是本研究首次报道的新 SNP。双荧光素酶报告基因检测系统的启动子检测结果表明，与野生型 GG（3.24±0.103）相比，CC 基因型的转录活性最强（13.67±0.578）。另一方面，研究结果表明，所选 microRNA（dme-miR-2279-5p）之一与 Hsp90AA1 3'-UTR 序列相互作用，并在等位基因 G 存在的情况下显著抑制报告基因活性（2.480±0.136）。在应激季节，携带突变型等位基因 C 的奶牛的 Hsp90AA1 表达量较高（4.18±0.928），而携带野生型等位基因 G 的奶牛的表达量较低（1.008±0.0.129）。总之，Hsp90AA1 基因的遗传变异与耐热性有关。这种关联可作为中国荷斯坦奶牛耐热性遗传选择的标记。
