The exon 29 c.3535A>T in the alpha-2-macroglobulin gene causing aberrant splice variants is associated with mastitis in dairy cattle
The exon 29 c.3535A>T in the alpha-2-macroglobulin gene causing aberrant splice variants is associated with mastitis in dairy cattle
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DOI：
10.1007/s00251-012-0639-8
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发表时间：
2012-08
期刊：
影响因子：
3.2
通讯作者：
Xiuge Wang;Jinming Huang;Lihong Zhao;Changfa Wang;Z. Ju;Qiuling Li;Chao Qi;Yan Zhang;Zebin Zhang;Wei Zhang;M. Hou;Jinduo Yuan;J. Zhong
中科院分区：
文献类型：
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作者：
Xiuge Wang;Jinming Huang;Lihong Zhao;Changfa Wang;Z. Ju;Qiuling Li;Chao Qi;Yan Zhang;Zebin Zhang;Wei Zhang;M. Hou;Jinduo Yuan;J. Zhong
Alpha-2-巨球蛋白 (A2M) 与蛋白酶结合，从而充当脊椎动物和无脊椎动物血浆和组织中病原体的防御屏障。采用实时定量聚合酶链反应（PCR）和同量异位标签相对和绝对定量方法测定乳腺炎感染乳腺组织中A2M mRNA和蛋白的表达水平。乳腺炎感染的乳腺组织中 A2M mRNA 和蛋白的表达显着高于健康组织。我们还使用逆转录 PCR 结合克隆测序在牛乳腺组织中鉴定了 23 个新型 A2Msplice 变体。这些剪接变体主要影响蛋白质的诱饵区、抑制区和硫酯区，这些区域在抑制病原体的蛋白酶方面发挥着关键的功能作用。基因组测序分析揭示了外显子 29 中的非同义 c.3535A>T 单核苷酸多态性 (SNP)，其位于推定的外显子剪接增强子内，可能是 A2M 基因产生异常剪接变体 A2M-AS4 的原因。我们的研究结果表明A2M基因可以通过选择性剪接机制发挥作用，可能对乳腺炎具有重要意义。这项研究为更好地了解牛 A2M 基因的功能以及外显子 SNP 对异常剪接变体产生的影响提供了线索。
Alpha-2-macroglobulin (A2M) binds proteases, thereby acting as defense barriers against pathogens in the plasma and tissues of vertebrates and invertebrates. Quantitative real-time polymerase chain reaction (PCR) and the isobaric tags for relative and absolute quantitation method were used to determine the expression levels of A2M mRNA and proteins in mastitis-infected mammary tissues. A2M mRNA and protein expression were significantly higher in mastitis-infected mammary tissues than those in healthy tissues. We also identified 23 novelA2Msplice variants in the bovine mammary tissues using reverse transcription PCR combined with clone sequencing. These splice variants predominantly affected the bait region, the inhibitory region, and the thioester region of the protein, which have the functional key roles in inhibiting the proteases of pathogens. Genomic sequencing analysis revealed a nonsynonymous c.3535A>T single-nucleotide polymorphism (SNP) in exon 29, which is located within a putative exonic splice enhancer and may be the reason why theA2Mgene produces the aberrant splice variantA2M-AS4. Our findings suggest that theA2Mgene can play its role by alternative splicing mechanism and it may be of significance against mastitis. This study provides clues to better understand the function of the bovineA2Mgene and the effects of the exonic SNP on the production of aberrant splice variants.