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首页> 外文期刊>Biochemical Genetics >Tissue-Specific Monoallelic Expression of Bovine AXL is Associated with DNA Methylation of Promoter DMR
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Tissue-Specific Monoallelic Expression of Bovine AXL is Associated with DNA Methylation of Promoter DMR

机译：牛AXL的组织特异性单相连表达与启动子DMR的DNA甲基化有关

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摘要

The AXL protein is a receptor tyrosine kinase and is often implicated in proliferation, migration and therapy resistance in various cancers. The AXL gene in humans is maternally expressed and paternally imprinted with differentially methylated regions (DMR) surrounding the promoter region. However, the imprinting status and epigenetic regulation of AXL gene in cattle remain unclear. Therefore, we explored the molecular structure along with the patterns of allelic expression and DNA methylation of the bovine AXL gene. First, the complete cDNA sequence of bovine AXL was gathered by Sanger method, from transcripts obtained from RT-PCR, 5 ' and 3 ' -RACE. In silico BLAST alignments showed that the longest mRNA sequence of bovine AXL consists of 19 exons and encodes a protein of 887 amino acids. We further analyzed the allelic expression of bovine AXL by employing single-nucleotide polymorphism (SNP)-based sequencing method. A SNP site (GenBank Accession no: rs210020651) found in exon 7 allowed us to distinguish the two parental alleles. Monoallelic expression of AXL was observed in four adult bovine tissues (heart, liver, spleen and fat), while biallelic expression was found in the other adult tissues such as the lung, kidney, muscle, brain and placenta. To determine whether the DNA methylation played a role in the tissue-specific imprinting of bovine AXL, we performed bisulfite sequencing of two regions: region 1 was a CpG island (CGI) in AXL promoter, mapping to 643 bp upstream of the transcription start site of AXL 5 '-v1 transcripts, while region two was homologous to the region of human AXL DMR, with 10 CpG sites overlapping the first translation start site (TSS1) of bovine AXL. In region 2, DNA from both monoallelic and biallelic expressed tissues were mostly found to be completely unmethylated. However, tissue-specific differential methylation patterns were found in monoallelic expressed tissues such as the heart and liver while hypomethylation was noted in the promoter CpG island in biallelic expressed tissues such as the lung. These observations demonstrated that the tissue-specific monoallelic expression of bovine AXL is dependent on the DNA methylation of its promoter region.
机译：AXL蛋白是受体酪氨酸激酶，通常涉及各种癌症的增殖，迁移和治疗抗性。人体中的AXL基因均用围绕启动子区周围的差异甲基化区域（DMR）致表达和伴随物质。然而，牛中AXL基因的印迹状态和表观遗传调节仍不清楚。因此，我们探讨了分子结构以及牛AXL基因的等位基因表达和DNA甲基化的模式。首先，通过Sanger方法从RT-PCR，5'和3'-RACE获得的转录物中收集完整的牛AXL的CDNA序列。在Silico Blast比赛中，牛AXL的最长mRNA序列由19个外显子组成，并编码887个氨基酸的蛋白质。我们通过采用单核苷酸多态性（SNP）的测序方法进一步分析了牛AXL的等位基因表达。在外显子7中发现的SNP网站（GenBank登录号：RS210020651）允许我们区分两个父母等位基因。在四个成人牛组织（心脏，肝脏，脾脏和脂肪）中观察到单独的AXL表达，而在其他成年组织如肺，肾，肌肉，脑和胎盘中发现双腿表达。为了确定DNA甲基化是否在牛AXL的组织特异性印迹中起作用的作用，我们对两种区域进行了亚硫酸硫酸氢盐测序：区域1是AXL启动子的CPG岛（CGI），在转录开始现场上游映射到643bp AXL 5'-V1转录物，而区域2对人AXL DMR区域同源，其中10个CPG位点重叠牛AXL的第一翻译开始点（TSS1）。在区域2中，大多数单独和双曲线表达组织的DNA是完全未甲基化的。然而，在单腹表达组织中发现组织特异性差异甲基化模式，例如心脏和肝脏，而在双雷梅尔的促进剂CpG岛中注意到如肺的促进剂CpG岛。这些观察结果表明，牛AXL的组织特异性单相连表达取决于其启动子区的DNA甲基化。

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  • 来源
    《Biochemical Genetics》 |2019年第6期|共12页
  • 作者

    Xu Da; Wang Guannan; Li Junliang; Zhang Cui; Chen Weina; Li Dongjie; Li Shijie;

  • 作者单位

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

    Hebei Univ Sci &

    Technol Coll Biosci &

    Bioengn Shijiazhuang 050018 Hebei Peoples R China;

    Agr Univ Hebei Coll Life Sci Baoding 071001 Peoples R China;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 遗传学;
  • 关键词

    Tissue-specific; DMR; AXL; Bovine;

    机译：组织特异性;DMR;AXL;牛;

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