Details

Autor(en) / Beteiligte

• Barsyte-Lovejoy, Dalia
• Li, Fengling
• Oudhoff, Menno J.
• Tatlock, John H.
• Dong, Aiping
• Zeng, Hong
• Wu, Hong
• Freeman, Spencer A.
• Schapira, Matthieu
• Senisterra, Guillermo A.
• Kuznetsova, Ekaterina
• Marcellus, Richard
• Allali-Hassani, Abdellah
• Kennedy, Steven
• Lambert, Jean-Philippe
• Couzens, Amber L.
• Aman, Ahmed
• Gingras, Anne-Claude
• Al-Awar, Rima
• Fish, Paul V.
• Gerstenberger, Brian S.
• Roberts, Lee
• Benn, Caroline L.
• Grimley, Rachel L.
• Braam, Mitchell J. S.
• Rossi, Fabio M. V.
• Sudol, Marius
• Brown, Peter J.
• Bunnage, Mark E.
• Owen, Dafydd R.
• Zaph, Colby
• Vedadi, Masoud
• Arrowsmith, Cheryl H.

Titel

(R)-PFI-2 is a potent and selective inhibitor of SETD7 methyltransferase activity in cells

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 2014-09, Vol.111 (35), p.12853-12858

Ort / Verlag

United States: National Academy of Sciences

Erscheinungsjahr

2014

Quelle

MEDLINE

Beschreibungen/Notizen

• SET domain containing (lysine methyltransferase) 7 (SETD7) is implicated in multiple signaling and disease related pathways with a broad diversity of reported substrates. Here, we report the discovery of (R)-PFI-2—a first-in-class, potent (K ᵢᵃᵖᵖ = 0.33 nM), selective, and cell-active inhibitor of the methyltransferase activity of human SETD7—and its 500-fold less active enantiomer, (S)-PFI-2. (R)-PFI-2 exhibits an unusual cofactor-dependent and substrate-competitive inhibitory mechanism by occupying the substrate peptide binding groove of SETD7, including the catalytic lysine-binding channel, and by making direct contact with the donor methyl group of the cofactor, S -adenosylmethionine. Chemoproteomics experiments using a biotinylated derivative of (R)-PFI-2 demonstrated dose-dependent competition for binding to endogenous SETD7 in MCF7 cells pretreated with (R)-PFI-2. In murine embryonic fibroblasts, (R)-PFI-2 treatment phenocopied the effects of Setd7 deficiency on Hippo pathway signaling, via modulation of the transcriptional coactivator Yes-associated protein (YAP) and regulation of YAP target genes. In confluent MCF7 cells, (R)-PFI-2 rapidly altered YAP localization, suggesting continuous and dynamic regulation of YAP by the methyltransferase activity of SETD7. These data establish (R)-PFI-2 and related compounds as a valuable tool-kit for the study of the diverse roles of SETD7 in cells and further validate protein methyltransferases as a druggable target class.