Department of Chemistry and ‡Department of Biochemistry and Molecular Biology, Colorado State University , Fort Collins, Colorado 80523, United States.
Biochemistry. 2013 Jul 16;52(28):4745-7. doi: 10.1021/bi400801q. Epub 2013 Jul 1.
U1A binds U1hpII, a hairpin RNA with a 10-nucleotide loop. A U1A mutant (ΔK50ΔM51) binds U1hpII-derived hairpins with shorter loops, making it an interesting scaffold for engineering or evolving proteins that bind similarly sized disease-related hairpin RNAs. However, a more detailed understanding of complexes involving ΔK50ΔM51 is likely a prerequisite to generating such proteins. Toward this end, we measured mutational effects for complexes involving U1A ΔK50ΔM51 and U1hpII-derived hairpin RNAs with seven- or eight-nucleotide loops and identified contacts that are critical to the stabilization of these complexes. Our data provide valuable insight into sequence-selective recognition of seven- or eight-nucleotide loop hairpins by an engineered RNA binding protein.
U1A 结合 U1hpII，这是一种具有 10 个核苷酸环的发夹 RNA。U1A 突变体（ΔK50ΔM51）结合具有较短环的 U1hpII 衍生发夹，使其成为工程或进化结合类似大小疾病相关发夹 RNA 的蛋白质的有趣支架。然而，更详细地了解涉及 ΔK50ΔM51 的复合物可能是生成此类蛋白质的前提。为此，我们测量了涉及 U1A ΔK50ΔM51 和 U1hpII 衍生的具有七或八核苷酸环的发夹 RNA 的复合物的突变效应，并确定了对这些复合物稳定至关重要的接触。我们的数据为工程 RNA 结合蛋白对七或八核苷酸环发夹的序列选择性识别提供了有价值的见解。
