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在翻译报告员中使用酶活性发现的DDX3X敏感性的决定因素

Kevin C Wilkins^1,2, Till Schroeder^1,3, Sohyun Gu¹

¹Department of Cell and Tissue Biology, University of California, San Francisco, San Francisco, California, 94143, USA.

bioRxiv : the preprint server for biology

|September 25, 2023

概括

翻译的螺旋酶活动报告器 (HART) 测量DDX3X活动. 这显示了DDX3X.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
生物化学生物化学

背景情况:

DDX3X是mRNA翻译的关键调节者.
复杂的5'未翻译区域 (5' UTR) 对于翻译控制至关重要.
了解DDX3X的机制对于细胞过程至关重要.

研究的目的:

开发一种用于测量DDX3X介导翻译的新型记者试验.
为了研究DDX3X依赖翻译的结构基础.
为了确定DDX3X的关键功能域.

主要方法:

开发用于翻译 (HART) 试验的酶活性报告器.
SHAPE-MaP (通过原料扩展和突变分析分析的选择性2'-基化) 来绘制RNA二次结构的地图.
对于HART报告员细胞表达的lentiviral输送.

主要成果:

哈特成功地测量了DDX3X依赖的翻译活动.
RNA结构的扰动会影响DDX3X的敏感性.
DDX3X的残留物38-44涉及到翻译机械相互作用.
DDX3X的核糖体关联和酶活性对其功能至关重要.

结论:

在核糖体扫描过程中,DDX3X通过与翻译机器相互作用来调节翻译.
哈特报告器是研究DDX3X依赖翻译的强大工具.
5' UTR 的结构特征对于 DDX3X 调节至关重要.

关键词:

这是一个DDX3XX.死亡盒螺旋酶是什么?在RNA中,有RNA基酶.核糖体扫描核糖体扫描开始翻译开始翻译它们是mRNARNA.

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