Identification of DYRK1B as a substrate of ERK1/2 and characterisation of the kinase activity of DYRK1B mutants from cancer and metabolic syndrome.
The dual-specificity tyrosine-phosphorylation-regulated kinase, DYRK1B, is expressed de novo during myogenesis, amplified or mutated in certain cancers and mutated in familial cases of metabolic syndrome. DYRK1B is activated by cis auto-phosphorylation on tyrosine-273 (Y273) within the activation loop during translation but few other DYRK1B phosphorylation sites have been characterised to date. Here, we demonstrate that DYRK1B also undergoes trans-autophosphorylation on serine-421 (S421) in vitro and in cells and that this site contributes to DYRK1B kinase activity. Whilst a DYRK1B(S421A) mutant was completely defective for p-S421 in cells, DYRK1B inhibitors caused only a partial loss of p-S421 suggesting the existence of an additional kinase that could also phosphorylate DYRK1B S421. Indeed, a catalytically inactive DYRK1B(D239A) mutant exhibited very low levels of p-S421 in cells but this was increased by KRAS(G12V). In addition, selective activation of the RAF-MEK1/2-ERK1/2 signalling pathway rapidly increased p-S421 in cells whereas activation of the stress kinases JNK or p38 could not. S421 resides within a Ser-Pro phosphoacceptor motif that is typical for ERK1/2 and recombinant ERK2 phosphorylated DYRK1B at S421 in vitro. Our results show that DYRK1B is a novel ERK2 substrate, uncovering new links between two kinases involved in cell fate decisions. Finally, we show that DYRK1B mutants that have recently been described in cancer and metabolic syndrome exhibit normal or reduced intrinsic kinase activity.
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK. anne.ashford@babraham.ac.uk.
Anne L
AL
Ashford
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.|Roche Innovation Center Basel, Basel, Switzerland.
Tom P J
TP
Dunkley
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.|Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
Mark
M
Cockerill
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.
Rachel A
RA
Rowlinson
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.
Lisa M
LM
Baak
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.
Raffaella
R
Gallo
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.
Kathryn
K
Balmanno
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.
Louise M
LM
Goodwin
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.
Richard A
RA
Ward
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK.
Pamela A
PA
Lochhead
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.|AstraZeneca, Waltham, MA, 02451, USA.
Sylvie
S
Guichard
AstraZeneca, Alderley Park, Macclesfield, Cheshire, SK10 4TG, UK.
Kevin
K
Hudson
Signalling Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, CB22 3AT, UK. simon.cook@babraham.ac.uk.
Simon J
SJ
Cook
D057809:HEK293 Cells; D006801:Humans; D024821:Metabolic Syndrome / Q000235:genetics* / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism*; D000097603:Dyrk Kinases
D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D000097603:Dyrk Kinases
DYRK1B; ERK1/2; KRAS; Phosphorylation; Protein kinase; RAF