Abstract
The mechanisms by which JAK2 is activated by the prevalent pseudokinase (JH2) V617F mutation in blood cancers remain elusive. Via structure-guided mutagenesis and transcriptional and functional assays, we identify a community of residues from the JH2 helix αC, SH2-JH2 linker and JH1 kinase domain that mediate V617F-induced activation. This circuit is broken by altering the charge of residues along the solvent-exposed face of the JH2 αC, which is predicted to interact with the SH2-JH2 linker and JH1. Mutations that remove negative charges or add positive charges, such as E596A/R, do not alter the JH2 V617F fold, as shown by the crystal structure of JH2 V617F E596A. Instead, they prevent kinase domain activation via modulation of the C-terminal residues of the SH2-JH2 linker. These results suggest strategies for selective V617F JAK2 inhibition, with preservation of wild-type function.
Original language | English (US) |
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Pages (from-to) | 1579-1591 |
Number of pages | 13 |
Journal | Biochemical Journal |
Volume | 473 |
Issue number | 11 |
DOIs | |
State | Published - Jun 1 2016 |
Keywords
- Inhibitor
- JAK-STAT signalling
- JAK2 V617F
- Kinase
- Myeloproliferative neoplasm (MPN)
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology