Univ. Bordeaux, Inserm, CNRS, ARNA Laboratory, U1212, UMR 5320, F-33000, Bordeaux, France.
Univ. Bordeaux, Inserm, CNRS, ARNA Laboratory, U1212, UMR 5320, Institut Européen de Chimie et Biologie, F-33600, Pessac, France.
Biomol NMR Assign. 2023 Jun;17(1):43-48. doi: 10.1007/s12104-023-10118-6. Epub 2023 Feb 1.
The initial pre-mRNA transcript in eukaryotes is processed by a large multi-protein complex in order to correctly cleave the 3' end, and to subsequently add the polyadenosine tail. This cleavage and polyadenylation specificity factor (CPSF) is composed of separate subunits, with structural information available for both isolated subunits and also larger assembled complexes. Nevertheless, certain key components of CPSF still lack high-resolution atomic data. One such region is the heterodimer formed between the first and second C-terminal domains of the endonuclease CPSF73, with those from the catalytically inactive CPSF100. Here we report the backbone and sidechain resonance assignments of a minimal C-terminal heterodimer of CPSF73-CPSF100 derived from the parasite Encephalitozoon cuniculi. The assignment process used several amino-acid specific labeling strategies, and the chemical shift values allow for secondary structure prediction.
真核生物中初始的前体 mRNA 转录本通过一个大型多蛋白复合物进行加工，以正确切割 3' 末端，并随后添加聚腺苷酸尾。这种切割和聚腺苷酸化特异性因子 (CPSF) 由独立的亚基组成，既有分离亚基的结构信息，也有更大组装复合物的结构信息。然而，CPSF 的某些关键成分仍然缺乏高分辨率的原子数据。其中一个区域是内切酶 CPSF73 的第一个和第二个 C 末端结构域与无催化活性的 CPSF100 之间形成的异二聚体。在这里，我们报告了来自寄生虫 Encephalitozoon cuniculi 的最小 C 末端 CPSF73-CPSF100 异二聚体的骨架和侧链共振分配。分配过程使用了几种氨基酸特异性标记策略，并且化学位移值允许进行二级结构预测。
