Substrate displacement of CK1 C-termini regulates kinase specificity

Sci Adv. 2024 May 10;10(19):eadj5185. doi: 10.1126/sciadv.adj5185. Epub 2024 May 10.


CK1 kinases participate in many signaling pathways, and their regulation is of meaningful biological consequence. CK1s autophosphorylate their C-terminal noncatalytic tails, and eliminating these tails increases substrate phosphorylation in vitro, suggesting that the autophosphorylated C-termini act as inhibitory pseudosubstrates. To test this prediction, we comprehensively identified the autophosphorylation sites on Schizosaccharomyces pombe Hhp1 and human CK1ε. Phosphoablating mutations increased Hhp1 and CK1ε activity toward substrates. Peptides corresponding to the C-termini interacted with the kinase domains only when phosphorylated, and substrates competitively inhibited binding of the autophosphorylated tails to the substrate binding grooves. Tail autophosphorylation influenced the catalytic efficiency with which CK1s targeted different substrates, and truncating the tail of CK1δ broadened its linear peptide substrate motif, indicating that tails contribute to substrate specificity as well. Considering autophosphorylation of both T220 in the catalytic domain and C-terminal sites, we propose a displacement specificity model to describe how autophosphorylation modulates substrate specificity for the CK1 family.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Casein Kinase 1 epsilon / genetics
  • Casein Kinase 1 epsilon / metabolism
  • Catalytic Domain
  • Humans
  • Mutation
  • Peptides / chemistry
  • Peptides / metabolism
  • Phosphorylation
  • Protein Binding
  • Schizosaccharomyces pombe Proteins* / chemistry
  • Schizosaccharomyces pombe Proteins* / genetics
  • Schizosaccharomyces pombe Proteins* / metabolism
  • Schizosaccharomyces* / genetics
  • Schizosaccharomyces* / metabolism
  • Substrate Specificity


  • Schizosaccharomyces pombe Proteins
  • Peptides
  • Casein Kinase 1 epsilon