3DXB

Structure of the UHM domain of Puf60 fused to thioredoxin

3DXB の概要

• エントリーDOI: 10.2210/pdb3dxb/pdb
• 分子名称: thioredoxin N-terminally fused to Puf60(UHM), CHLORIDE ION, 1,2-ETHANEDIOL, ... (4 entities in total)
• 機能のキーワード: splicing, fbp interacting repressor, uhm, rrm, electron transport, redox-active center, transport, transcription
• 由来する生物種: Escherichia coli O157:H7; 詳細
• 細胞内の位置: Nucleus : Q9UHX1
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 197579.32

構造登録者

Corsini, L.,Hothorn, M.,Scheffzek, K.,Stier, G.,Sattler, M. (登録日: 2008-07-24, 公開日: 2008-10-28, 最終更新日: 2024-10-30)

主引用文献

Corsini, L.,Hothorn, M.,Stier, G.,Rybin, V.,Scheffzek, K.,Gibson, T.J.,Sattler, M.
Dimerization and Protein Binding Specificity of the U2AF Homology Motif of the Splicing Factor Puf60.
J.Biol.Chem., 284:630-639, 2009

PubMed Abstract: PUF60 is an essential splicing factor functionally related and homologous to U2AF(65). Its C-terminal domain belongs to the family of U2AF (U2 auxiliary factor) homology motifs (UHM), a subgroup of RNA recognition motifs that bind to tryptophan-containing linear peptide motifs (UHM ligand motifs, ULMs) in several nuclear proteins. Here, we show that the Puf60 UHM is mainly monomeric in physiological buffer, whereas its dimerization is induced upon the addition of SDS. The crystal structure of PUF60-UHM at 2.2 angstroms resolution, NMR data, and mutational analysis reveal that the dimer interface is mediated by electrostatic interactions involving a flexible loop. Using glutathione S-transferase pulldown experiments, isothermal titration calorimetry, and NMR titrations, we find that Puf60-UHM binds to ULM sequences in the splicing factors SF1, U2AF65, and SF3b155. Compared with U2AF65-UHM, Puf60-UHM has distinct binding preferences to ULMs in the N terminus of SF3b155. Our data suggest that the functional cooperativity between U2AF65 and Puf60 may involve simultaneous interactions of the two proteins with SF3b155.

PubMed: 18974054
DOI: 10.1074/jbc.M805395200

実験手法

X-RAY DIFFRACTION (2.2 Å)