Cencini Aura, Rilievo Graziano, Cecconello Alessandro, Tonolo Federica, Babbucci Massimiliano, Negrisolo Enrico, Magro Massimiliano, Vianello Fabio
Department of Comparative Biomedicine and Food Science (BCA), University of Padova, Campus Agripolis, viale dell'Università 16, 35020 Legnaro (PD), Italy.
Department of Molecular and Translational Medicine (DMMT), University of Brescia, viale Europa 11, 25135 Brescia (BS), Italy.
Nucleic Acids Res. 2025 May 22;53(10). doi: 10.1093/nar/gkaf429.
Non-coding RNAs belong to a heterogenous family that, among other functions, acts as a biomolecular regulator of gene expression. In particular, lncRNAs, which are estimated to be as numerous as coding RNAs in humans, are thought to interact with genomic DNA to form triple helices. However, experimental evidence of their involvement with processes, such as chromatin structure dynamics or RNA transcription, is still missing. Here, a mechanism of transcription enhancement/inhibition is described, where hybrid RNA-DNA triplexes regulate transcription rates in Escherichia coli promoter-based designed architectures. Sequences associated with triplexes were identified in a library of bacterial promoters and characterized in vitro, followed by a synthetic biology approach to verify their ability to control transcription and translation. A model of the triplex-promoter complex was produced showing that transcription enhancement is due to a distortion of the duplex DNA as a consequence of its conjugation with RNA in the triplex assembly. These results point at a mechanism of RNA function that is still unknown and could be common in more complex organisms, such as metazoans including mammals, where non-coding RNAs are more abundant and are believed to play a fundamental role in determining hetero/euchromatin and transcription modulation.
非编码RNA属于一个异质家族,除其他功能外,它还作为基因表达的生物分子调节剂发挥作用。特别是长链非编码RNA(lncRNA),据估计其数量与人类中的编码RNA一样多,被认为与基因组DNA相互作用形成三链螺旋。然而,关于它们参与染色质结构动态变化或RNA转录等过程的实验证据仍然缺乏。在此,我们描述了一种转录增强/抑制机制,其中RNA-DNA杂交三链体在基于大肠杆菌启动子设计的结构中调节转录速率。在细菌启动子文库中鉴定出与三链体相关的序列,并在体外进行了表征,随后采用合成生物学方法验证它们控制转录和翻译的能力。构建了三链体-启动子复合物模型,结果表明转录增强是由于双链DNA在三链体组装过程中与RNA结合导致其扭曲所致。这些结果指向一种未知的RNA功能机制,这种机制可能在更复杂的生物体中普遍存在,比如包括哺乳动物在内的后生动物,在这些生物体中,非编码RNA更为丰富,并且被认为在决定异染色质/常染色质和转录调控方面发挥着重要作用。