Hedaya Omar M, Subbaiah Kadiam C Venkata, Jiang Feng, Xie Li Huitong, Wu Jiangbin, Khor EngSoon, Zhu Mingyi, Mathews David H, Proschel Chris, Yao Peng
Aab Cardiovascular Research Institute, Department of Medicine, University of Rochester School of Medicine & Dentistry, Rochester, New York 14642.
Department of Biochemistry & Biophysics, University of Rochester School of Medicine & Dentistry, Rochester, New York 14642.
bioRxiv. 2023 Jun 15:2023.06.15.545153. doi: 10.1101/2023.06.15.545153.
Translation of upstream open reading frames (uORFs) typically abrogates translation of main (m)ORFs. The molecular mechanism of uORF regulation in cells is not well understood. Here, we identified a double-stranded RNA (dsRNA) structure residing within the uORF that augments uORF translation and inhibits mORF translation. Antisense oligonucleotides (ASOs) that disrupt this dsRNA structure promote mORF translation, while ASOs that base-pair immediately downstream (i.e., forming a bimolecular double-stranded region) of either the uORF or mORF start codon enhance uORF or mORF translation, respectively. Human cardiomyocytes and mice treated with a uORF-enhancing ASO showed reduced cardiac GATA4 protein levels and increased resistance to cardiomyocyte hypertrophy. We further show the general utility of uORF-dsRNA- or mORF- targeting ASO to regulate mORF translation for other mRNAs. Our work demonstrates a regulatory paradigm that controls translational efficiency and a useful strategy to alter protein expression and cellular phenotypes by targeting or generating dsRNA downstream of a uORF or mORF start codon.
dsRNA within uORF activates uORF translation and inhibits mORF translation. ASOs that target the dsRNA can either inhibit or enhance mORF translation. ASOs can be used to impede hypertrophy in human cardiomyocytes and mouse hearts.uORF- and mORF-targeting ASOs can be used to control translation of multiple mRNAs.
上游开放阅读框（uORF）的翻译通常会消除主要（m）开放阅读框的翻译。细胞中uORF调控的分子机制尚不清楚。在这里，我们在uORF中鉴定出一种双链RNA（dsRNA）结构，它增强了uORF的翻译并抑制了mORF的翻译。破坏这种dsRNA结构的反义寡核苷酸（ASO）可促进mORF的翻译，而在uORF或mORF起始密码子下游立即碱基配对（即形成双分子双链区域）的ASO分别增强uORF或mORF的翻译。用增强uORF的ASO处理的人类心肌细胞和小鼠显示心脏GATA4蛋白水平降低，对心肌细胞肥大的抵抗力增加。我们进一步证明了靶向uORF-dsRNA或mORF的ASO在调节其他mRNA的mORF翻译方面的普遍效用。我们的工作展示了一种控制翻译效率的调控模式，以及一种通过靶向uORF或mORF起始密码子下游的dsRNA或产生dsRNA来改变蛋白质表达和细胞表型的有用策略。
uORF内的dsRNA激活uORF翻译并抑制mORF翻译。靶向dsRNA的ASO可以抑制或增强mORF翻译。ASO可用于阻止人类心肌细胞和小鼠心脏的肥大。靶向uORF和mORF的ASO可用于控制多种mRNA的翻译。
