4TU7

Structure of U2AF65 D231V variant with BrU5 DNA

4TU7 の概要

• エントリーDOI: 10.2210/pdb4tu7/pdb
• 関連するPDBエントリー: 4TU8 4TU9
• 分子名称: Splicing factor U2AF 65 kDa subunit, DNA (5'-D(*UP*UP*UP*UP*(BRU)P*UP*U)-3'), 1,4-DIETHYLENE DIOXIDE, ... (7 entities in total)
• 機能のキーワード: pre-mrna splicing factor, rna recognition motif, rna binding protein-dna complex, rna binding protein/dna
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Nucleus: P26368
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 44409.04

構造登録者

Agrawal, A.A.,Jenkins, J.L.,Kielkopf, C.L. (登録日: 2014-06-24, 公開日: 2014-11-26, 最終更新日: 2023-12-27)

主引用文献

Agrawal, A.A.,McLaughlin, K.J.,Jenkins, J.L.,Kielkopf, C.L.
Structure-guided U2AF65 variant improves recognition and splicing of a defective pre-mRNA.
Proc.Natl.Acad.Sci.USA, 111:17420-17425, 2014

PubMed Abstract: Purine interruptions of polypyrimidine (Py) tract splice site signals contribute to human genetic diseases. The essential splicing factor U2AF(65) normally recognizes a Py tract consensus sequence preceding the major class of 3' splice sites. We found that neurofibromatosis- or retinitis pigmentosa-causing mutations in the 5' regions of Py tracts severely reduce U2AF(65) affinity. Conversely, we identified a preferred binding site of U2AF(65) for purine substitutions in the 3' regions of Py tracts. Based on a comparison of new U2AF(65) structures bound to either A- or G-containing Py tracts with previously identified pyrimidine-containing structures, we expected to find that a D231V amino acid change in U2AF(65) would specify U over other nucleotides. We found that the crystal structure of the U2AF(65)-D231V variant confirms favorable packing between the engineered valine and a target uracil base. The D231V amino acid change restores U2AF(65) affinity for two mutated splice sites that cause human genetic diseases and successfully promotes splicing of a defective retinitis pigmentosa-causing transcript. We conclude that reduced U2AF(65) binding is a molecular consequence of disease-relevant mutations, and that a structure-guided U2AF(65) variant is capable of manipulating gene expression in eukaryotic cells.

PubMed: 25422459
DOI: 10.1073/pnas.1412743111

実験手法

X-RAY DIFFRACTION (2.087 Å)