6SLO

Crystal structure of PUF60 UHM domain in complex with 7,8 dimethoxyperphenazine

6SLO の概要

• エントリーDOI: 10.2210/pdb6slo/pdb
• 分子名称: Thioredoxin,Poly(U)-binding-splicing factor PUF60, MAGNESIUM ION, 2-[4-[3-(8-chloranyl-2,3-dimethoxy-phenothiazin-10-yl)propyl]piperazin-1-yl]ethanol, ... (4 entities in total)
• 機能のキーワード: uhm domain, splicing inhibitor, splicing
• 由来する生物種: Citrobacter freundii MGH 56; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 99664.37

構造登録者

Jagtap, P.K.A.,Kubelka, T.,Bach, T.,Sattler, M. (登録日: 2019-08-20, 公開日: 2020-09-09, 最終更新日: 2024-10-09)

主引用文献

Jagtap, P.K.A.,Kubelka, T.,Soni, K.,Will, C.L.,Garg, D.,Sippel, C.,Kapp, T.G.,Potukuchi, H.K.,Schorpp, K.,Hadian, K.,Kessler, H.,Luhrmann, R.,Hausch, F.,Bach, T.,Sattler, M.
Identification of phenothiazine derivatives as UHM-binding inhibitors of early spliceosome assembly.
Nat Commun, 11:5621-5621, 2020

PubMed Abstract: Interactions between U2AF homology motifs (UHMs) and U2AF ligand motifs (ULMs) play a crucial role in early spliceosome assembly in eukaryotic gene regulation. UHM-ULM interactions mediate heterodimerization of the constitutive splicing factors U2AF65 and U2AF35 and between other splicing factors that regulate spliceosome assembly at the 3' splice site, where UHM domains of alternative splicing factors, such as SPF45 and PUF60, contribute to alternative splicing regulation. Here, we performed high-throughput screening using fluorescence polarization assays with hit validation by NMR and identified phenothiazines as general inhibitors of UHM-ULM interactions. NMR studies show that these compounds occupy the tryptophan binding pocket of UHM domains. Co-crystal structures of the inhibitors with the PUF60 UHM domain and medicinal chemistry provide structure-activity-relationships and reveal functional groups important for binding. These inhibitors inhibit early spliceosome assembly on pre-mRNA substrates in vitro. Our data show that spliceosome assembly can be inhibited by targeting UHM-ULM interactions by small molecules, thus extending the toolkit of splicing modulators for structural and biochemical studies of the spliceosome and splicing regulation.

PubMed: 33159082
DOI: 10.1038/s41467-020-19514-1

実験手法

X-RAY DIFFRACTION (1.94 Å)