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通过上游开放的阅读框架元素对小鼠皮蛋白表达的翻译控制

Kommireddy Vasu¹, Iyappan Ramachandiran¹, Aayushi Chechi¹

¹Department of Cardiovascular and Metabolic Sciences, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA.

|September 13, 2023

概括

在adiponectin (ADIPOQ) mRNA 5'-leader中的一个新的上游开放阅读框架 (uORF) 调节了它的翻译. 破坏这种uORF可以增强阿迪波内克丁的表达,并改善葡萄糖的吸收.

科学领域:

分子生物学分子生物学
代谢调节代谢调节代谢调节
控制基因表达控制基因表达

背景情况:

由ADIPOQ基因编码的阿迪波涅丁对胰岛素敏感性至关重要,其血水平通过后转录机制被抗糖尿病药物调节.
阿迪波涅丁mRNA的5'-领导者含有影响蛋白质翻译的调节元素,但它们的确切作用,特别是在压力下,仍然不清楚.

研究的目的:

为了研究上游开放阅读框架 (uORFs) 在阿迪波涅丁mRNA 5'-leader中的功能.
阐明这些uORFs在调节腺素表达和其下游代谢效应中的作用,特别是在应对细胞压力的过程中.

主要方法:

记者测试被用来分析阿迪波涅克丁mRNA5'-领导者的cis调节功能,包括潜在的uORFs的删除和突变.
综合应激反应 (ISR) 激活使用thapsigargin (TG) 和通过多元体分析进行翻译分析.
采用CRISPR-Cas9基因编辑来破坏差异化3T3-L1脂肪细胞中识别的uORF.

主要成果:

删除分析确定了5'-领导体中的抑制性cis-regulatory序列,其中包含两个uORF. 突变的uORF起始编码子增加了记者翻译大约3倍.
塔普西加林诱导的内等质网膜 (ER) 压力和ISR增强的记者表达,依赖eIF2α酸化,证实了uORF在压力中介翻译中的作用.
在脂肪细胞中,CRISPR介导的uORF中断显著增加了阿迪波涅丁的表达,三糖醇的积累和葡萄糖的吸收,同时抑制了邻近细胞中葡萄糖生成酶的表达.

结论:

这项研究在阿迪波涅克丁mRNA 5'-leader中确定了一个功能性uORF,它在调节阿迪波涅克丁翻译方面发挥着关键作用.
这种uORF调解了压力诱导的阿迪波涅丁的翻译控制,影响了脂肪细胞功能和细胞间代谢信号传递.
针对这种uORF为胰岛素抵抗等代谢障碍提供了潜在的治疗策略.

关键词:

阿迪波克 (Adipoq) 是一个克里斯普尔-Cas9是什么意思上游开放的阅读框架.脂肪细胞的脂肪细胞.没有任何的Adiponectin.单核细胞启动因子2alpha是什么综合应激反应综合应激反应翻译翻译翻译翻译翻译翻译

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