Gordon Blaine H, Silvers Robert
Department of Chemistry & Biochemistry, Florida State University, 95 Chieftan Way, Tallahassee, FL, 32306, USA.
Institute of Molecular Biophysics, Florida State University, 91 Chieftan Way, Tallahassee, FL, 32306, USA.
Biomol NMR Assign. 2025 Jun;19(1):165-173. doi: 10.1007/s12104-025-10232-7. Epub 2025 Apr 30.
Human La-related protein 6 (HsLARP6) participates in the post-transcriptional regulation of type I collagen biosynthesis and is involved in the onset and progression of fibroproliferative disease. The RNA-binding protein HsLARP6 recognizes a hairpin structure known as the 5' stem-loop (5'SL) located at the junction of 5' untranslated and coding regions of type I collagen mRNA. Despite extensive biochemical and functional studies of the interaction between HsLARP6 and the 5'SL motif, the lack of high-resolution molecular data significantly hampers our understanding of the binding mechanism. Here, we introduced a shorter 5'SL model, named A2M5, reducing the molecular size of the protein-RNA complex as well as spectral overlap in RNA-based spectra. Furthermore, we reported the near-complete backbone and side chain resonance assignment of the La domain of HsLARP6 in a 1:1 complex with the A2M5 model RNA. These results will provide a significant platform for future NMR spectroscopic studies of 5'SL binding to the La domain of HsLARP6.
人La相关蛋白6（HsLARP6）参与I型胶原蛋白生物合成的转录后调控，并与纤维增生性疾病的发生和发展有关。RNA结合蛋白HsLARP6识别一种发夹结构，称为5'茎环（5'SL），位于I型胶原蛋白mRNA的5'非翻译区和编码区的交界处。尽管对HsLARP6与5'SL基序之间的相互作用进行了广泛的生化和功能研究，但缺乏高分辨率分子数据严重阻碍了我们对结合机制的理解。在这里，我们引入了一个较短的5'SL模型，命名为A2M5，它减小了蛋白质-RNA复合物的分子大小以及基于RNA的光谱中的光谱重叠。此外，我们报道了HsLARP6的La结构域与A2M5模型RNA形成 的1:1复合物中几乎完整的主链和侧链共振归属。这些结果将为未来关于5'SL与HsLARP6的La结构域结合的核磁共振光谱研究提供一个重要平台。
