加勒-3与RNA没有直接相互作用
Egan L Peltan1,2, Nicholas M Riley2,3, Ryan A Flynn4,5
1Department of Chemical and Systems Biology, Stanford University School of Medicine, 269 Campus Drive CCSR 4145 Stanford, CA 94305, United States.
Glycobiology
|October 10, 2023
概括
加勒-3不是一种直接的RNA结合蛋白. 研究表明,分离RNA-蛋白交叉链的抗体是由于hnRNPA2B1,而不是galectin-3本身.
科学领域:
- 分子生物学分子生物学
- 葡萄糖生物学 葡萄糖生物学
- 在RNA生物学,RNA生物学.
背景情况:
- 已知一种糖甘结合蛋白的加勒-3被提出作为一种具有RNA结合能力的双重功能蛋白.
- 这种假定作用表明了糖生物学和RNA生物学之间的联系,特别是在mRNA前拼接中.
- 以前的证据依赖于缺乏遗传验证的亲和试剂,因此直接RNA相互作用未得到证实.
研究的目的:
- 研究加勒-3的直接RNA结合能力.
- 为了确定galectin-3是否在细胞过程中与RNA相互作用,如mRNA前成熟.
- 澄清之前观察到的涉及加勒-3的RNA-蛋白关联的分子基础.
主要方法:
- 使用抗体对抗内源性人体胆素-3用于RNA-蛋白交叉链接的免疫沉 (IP).
- 对IP分离物进行了蛋白质组分析.
- 产生了LGALS3淘汰细胞系和HNRNPA2B1淘汰细胞系.
- 引入了LGALS3的内源C终端位置的表位标签,以进行有针对性的隔离.
主要成果:
- 抗素-3抗体分离了RNA-蛋白交叉链接,但这是独立于LGALS3基因表达的.
- 蛋白质组分析显示,RNA结合蛋白 hnRNPA2B1 在分离物中非常丰富.
- 基因废除HNRNPA2B1,但不是LGALS3,废除了RNA-蛋白质交叉链路隔离.
- 隔离带表位标记的 galectin-3 并没有产生RNA-蛋白质交叉链接.
结论:
- 盖列-3似乎不会与RNA直接相互作用.
- 观察到的RNA关联很可能是由于涉及hNRNPA2B1.1的交叉反应或间接相互作用.
- 鉴定galectin-3为RNA结合蛋白可能是一个误解,特别是在HeLa细胞中.
- 建议使用基因删除和内源性表位标签来精确评估蛋白质-RNA相互作用.
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