PT  - JOURNAL ARTICLE
AU  - van de Kooij, Bert
AU  - Creixell, Pau
AU  - van Vlimmeren, Anne
AU  - Joughin, Brian A.
AU  - Miller, Chad J.
AU  - Haider, Nasir
AU  - Linding, Rune
AU  - Stambolic, Vuk
AU  - Turk, Benjamin E.
AU  - Yaffe, Michael B.
TI  - Comprehensive Substrate Specificity Profiling of the Human Nek Kinome Reveals Unexpected Signaling Outputs
AID  - 10.1101/515221
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 515221
4099  - http://biorxiv.org/content/early/2019/01/08/515221.short
4100  - http://biorxiv.org/content/early/2019/01/08/515221.full
AB  - Human NimA-related kinases (Neks) have multiple mitotic and non-mitotic functions, but few substrates are known. We systematically determined the phosphorylation-site motifs for the entire Nek kinase family, except for Nek11. While all Nek kinases strongly select for hydrophobic residues in the −3 position, the family separates into four distinct groups based on specificity for a serine versus threonine phospho-acceptor, and preference for basic or acidic residues in other positions. Unlike Nek1-Nek9, Nek10 is a dual-specificity kinase that efficiently phosphorylates itself and peptide substrates on serine and tyrosine, and its activity is enhanced by tyrosine auto-phosphorylation. Nek10 dual-specificity depends on residues in the HRD+2 and APE-4 positions that are uncommon in either serine/threonine or tyrosine kinases. Finally, we show that the phosphorylation-site motifs for the mitotic kinases Nek6, Nek7 and Nek9 are essentially identical to that of their upstream activator Plk1, suggesting that Nek6/7/9 function as phospho-motif amplifiers of Plk1 signaling.