Comprehensive analysis of translation from overexpressed circular RNAs reveals pervasive translation from linear transcripts.
Comprehensive analysis of translation from overexpressed circular RNAs reveals pervasive translation from linear transcripts.
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DOI：
10.1093/nar/gkaa704
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发表时间：
2020-10-09
影响因子：
14.9
通讯作者：
Meister G
中科院分区：
文献类型：
--
作者：
Ho-Xuan H;Glažar P;Latini C;Heizler K;Haase J;Hett R;Anders M;Weichmann F;Bruckmann A;Van den Berg D;Hüttelmaier S;Rajewsky N;Hackl C;Meister G
环状RNA（circRNA）是一类广泛存在的保守RNA，由下游5′剪接位点反向剪接至上游3′剪接位点而产生。CircRNA是组织特异性的，并且与包括癌症在内的疾病有关。例如，它们可以作为microRNA（miRNAs）或RNA结合蛋白（RBP）的海绵。此外，一些包含开放阅读框架（ORF），并可能被翻译。然而，这些肽的功能相关性在很大程度上仍然难以捉摸。在这里，我们报告了circZNF609的ORF在从circZNF609过表达构建体表达时被有效翻译。但未检测到内源性蛋白质。此外，circZNF 609翻译的起始不依赖于m6A生成酶胃L3或RNA序列元件，如内部核糖体进入位点（IRES）。令人惊讶的是，一项全面的突变分析显示，在产生circZNF 609方面有缺陷的缺失构建体仍然产生所观察到的蛋白质产物。这表明，明显的circZNF 609翻译起源于过表达质粒的反式剪接副产物，并强调需要仔细评估circRNA过表达构建体，特别是在进行功能研究时。
Circular RNAs (circRNAs) encompass a widespread and conserved class of RNAs, which are generated by back-splicing of downstream 5′ to upstream 3′ splice sites. CircRNAs are tissue-specific and have been implicated in diseases including cancer. They can function as sponges for microRNAs (miRNAs) or RNA binding proteins (RBPs), for example. Moreover, some contain open reading frames (ORFs) and might be translated. The functional relevance of such peptides, however, remains largely elusive. Here, we report that the ORF of circZNF609 is efficiently translated when expressed from a circZNF609 overexpression construct. However, endogenous proteins could not be detected. Moreover, initiation of circZNF609 translation is independent of m6A-generating enzyme METTL3 or RNA sequence elements such as internal ribosome entry sites (IRESs). Surprisingly, a comprehensive mutational analysis revealed that deletion constructs, which are deficient in producing circZNF609, still generate the observed protein products. This suggests that the apparent circZNF609 translation originates from trans-splicing by-products of the overexpression plasmids and underline that circRNA overexpression constructs need to be evaluated carefully, particularly when functional studies are performed.
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DOI：
10.1261/rna.053561.115
发表时间：
2016-06
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Carlevaro-Fita J;Rahim A;Guigó R;Vardy LA;Johnson R
通讯作者：
Johnson R
影响因子：
64.5
作者：
Kleaveland B;Shi CY;Stefano J;Bartel DP
通讯作者：
Bartel DP
影响因子：
14.9
作者：
Braun, S;Domdey, H;Wiebauer, K
通讯作者：
Wiebauer, K
影响因子：
4.8
作者：
COCQUERELLE, C;MASCREZ, B;BAILLEUL, B
通讯作者：
BAILLEUL, B
影响因子：
4.6
作者：
Gruner H;Cortés-López M;Cooper DA;Bauer M;Miura P
通讯作者：
Miura P