College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Gene. 2021 Jul 30;791:145710. doi: 10.1016/j.gene.2021.145710. Epub 2021 May 11.
Speckled 100 kDa (Sp100) plays an important role in the antiviral immune response, however, little is known about porcine Sp100. In this study, porcine Sp100 was cloned and its response to interferon (IFN) α was identified. We obtained the cDNA (V1) of the gene, SP100, and seven alternative splicing variants (V2-8). Isoform V1 encoded a 386 amino acid protein and contained a homogeneously-staining region (HSR) domain. Isoforms V3, 4, 6 and 7 were deletion/insertion variants and contained HSR domain as V1. The splicing of porcine SP100 was very complicated and many transcripts existed as revealed by cloning and minigene analyses. Using GFP-fusion constructs isoforms V1, 3, 4, 6 and 7 were localized to nucleus and the nuclear localization signal was identified as PSNRKRR at positions 331-337 of V1. Porcine SP100 was unevenly distributed in all tissues studied and differentially expressed between pigs with different disease-resistance/susceptibilities. Porcine SP100 was strongly increased by IFNα due to the existence of an IFN-stimulated response element in the promoter. A single nucleotide - 70A > C polymorphism enhanced promoter activity. The results provided the basis for determining the role of Sp100 in antiviral responses and may assist in breeding pigs with high disease-resistance.
斑驳蛋白 100kDa（Sp100）在抗病毒免疫反应中起着重要作用，但目前对猪 Sp100 的了解甚少。本研究克隆了猪 Sp100 并鉴定了其对干扰素（IFN）α的反应。我们获得了基因 SP100 的 cDNA（V1）和 7 个选择性剪接变体（V2-8）。异构体 V1 编码一个 386 个氨基酸的蛋白质，含有均匀染色区（HSR）结构域。异构体 V3、4、6 和 7 是缺失/插入变体，含有作为 V1 的 HSR 结构域。猪 SP100 的剪接非常复杂，通过克隆和小基因分析揭示了许多转录本的存在。使用 GFP 融合构建体，异构体 V1、3、4、6 和 7 定位于细胞核，核定位信号被鉴定为 V1 中 331-337 位的 PSNRKRR。猪 SP100 在所有研究的组织中分布不均匀，并且在具有不同疾病抗性/易感性的猪之间表达不同。由于启动子中存在干扰素刺激反应元件，猪 SP100 被 IFNα 强烈上调。单个核苷酸-70A>C 多态性增强了启动子活性。这些结果为确定 Sp100 在抗病毒反应中的作用提供了基础，并可能有助于培育具有高疾病抗性的猪。
