Title: Comprehensive substrate specificity profiling of the human Nek kinome reveals unexpected signaling outputs.

Authors: van de Kooij, Bert; Creixell, Pau; van Vlimmeren, Anne; Joughin, Brian A; Miller, Chad J; Haider, Nasir; Simpson, Craig D; Linding, Rune; Stambolic, Vuk; Turk, Benjamin E; Yaffe, Michael B

Published In Elife, (2019 05 24)

Abstract: 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.

PubMed ID: 31124786

MeSH Terms: Humans; NIMA-Related Kinases/chemistry; NIMA-Related Kinases/metabolism*; Phosphorylation; Serine/metabolism; Signal Transduction*; Substrate Specificity*; Threonine/metabolism