RT Journal Article
SR Electronic
T1 Investigating the determinants of mammalian Mastl kinase activation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.30.179580
DO 10.1101/2020.06.30.179580
A1 Mehmet Erguven
A1 M. Kasim Diril
YR 2020
UL http://biorxiv.org/content/early/2020/07/01/2020.06.30.179580.abstract
AB Mastl (Greatwall) kinase is an essential mitotic protein kinase. Mastl is an atypical member of AGC family with a unique long stretch of non-conserved middle region. The mechanism of its phosphorylation dependent activation has been studied in Xenopus egg extracts, revealing several phosphosites that were suggested to be crucial for kinase activation. These residues correspond to T193 and T206 in the activation loop, and S861 in the C-tail of mouse Mastl. By combining a chemically inducible knockout system to deplete the endogenous Mastl and a viral expression system to ectopically express the mutant variants, we obtained a viable knockout clone that expresses the S861A and S861D mutants. We observed that proliferation rates of the MastlS861A and MastlS861D clones were comparable. Our results have revealed that phosphorylation of the turn motif phosphosite (S861) is auxiliary and it is not indispensable for Mastl function.Competing Interest StatementThe authors have declared no competing interest.4-OHT4-Hydroxytamoxifen;DMSO;dimethyl sulfoxide;HAHuman influenza hemagglutinin;HFHydrophobic Motif;MEFMouse Embryonic Fibroblast;MUTMutant;NCMRNon-Conserved Middle Region;NLTN-Lobe Tether;PDBThe Protein Data Bank;SACSpindle Assembly Checkpoint;WTWild Type.