Jagtap, Pravin Kumar Ankush ; Kubelka, Tomáš ; Soni, Komal ; Will, Cindy L. ; Garg, Divita ; Sippel, Claudia ; Kapp, Tobias G. ; Potukuchi, Harish Kumar ; Schorpp, Kenji ; Hadian, Kamyar ; Kessler, Horst ; Lührmann, Reinhard ; Hausch, Felix ; Bach, Thorsten ; Sattler, Michael (2024)
Identification of phenothiazine derivatives as UHM-binding inhibitors of early spliceosome assembly.
In: Nature Communications, 2020, 11 (1)
doi: 10.26083/tuprints-00023981
Artikel, Zweitveröffentlichung, Verlagsversion
 | Es ist eine neuere Version dieses Eintrags verfügbar. |
Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2024 |
| Autor(en): | Jagtap, Pravin Kumar Ankush ; Kubelka, Tomáš ; Soni, Komal ; Will, Cindy L. ; Garg, Divita ; Sippel, Claudia ; Kapp, Tobias G. ; Potukuchi, Harish Kumar ; Schorpp, Kenji ; Hadian, Kamyar ; Kessler, Horst ; Lührmann, Reinhard ; Hausch, Felix ; Bach, Thorsten ; Sattler, Michael |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Identification of phenothiazine derivatives as UHM-binding inhibitors of early spliceosome assembly |
| Sprache: | Englisch |
| Publikationsjahr: | 25 September 2024 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 6 November 2020 |
| Ort der Erstveröffentlichung: | London |
| Verlag: | Springer Nature |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Nature Communications |
| Jahrgang/Volume einer Zeitschrift: | 11 |
| (Heft-)Nummer: | 1 |
| Kollation: | 11 Seiten |
| DOI: | 10.26083/tuprints-00023981 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/23981 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (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. |
| Freie Schlagworte: | Molecular modelling, NMR spectroscopy, RNA-binding proteins, Structural biology, X-ray crystallography |
| ID-Nummer: | Artikel-ID: 5621 |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-239811 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut |
| Hinterlegungsdatum: | 25 Sep 2024 11:46 |
| Letzte Änderung: | 26 Sep 2024 07:32 |
| PPN: | |
| Export: | |
| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
- Identification of phenothiazine derivatives as UHM-binding inhibitors of early spliceosome assembly. (deposited 25 Sep 2024 11:46) [Gegenwärtig angezeigt]
 |
Frage zum Eintrag |
Optionen (nur für Redakteure)
 |
Redaktionelle Details anzeigen |
Drucken
Impressum