4TU8

STRUCTURE OF U2AF65 VARIANT WITH BRU5A6 DNA

Summary for 4TU8

• Entry DOI: 10.2210/pdb4tu8/pdb
• Related: 4TU7 4TU9
• Descriptor: Splicing factor U2AF 65 kDa subunit, DNA (5'-D(*UP*UP*UP*UP*(BRU)P*DA*U)-3'), 1,4-DIETHYLENE DIOXIDE, ... (8 entities in total)
• Functional Keywords: rna splicing factor, rna recognition motif, rna binding protein, rna binding protein-dna complex, rna binding protein/dna
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 44745.49

Authors

MCLAUGHLIN, K.J.,JENKINS, J.L.,Agrawal, A.A.,KIELKOPF, C.L. (deposition date: 2014-06-24, release date: 2014-11-26, Last modification date: 2023-12-27)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.918 Å)