Hierarchical phosphorylation within the ankyrin repeat domain defines a phosphoregulatory loop that regulates notch transcriptional activity

Prathibha Ranganathan, Rodrigo Vasquez-Del Carpio, Fred M. Kaplan, Hong Wang, Ashu Gupta, Jeffrey D. VanWye, Anthony J Capobianco

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The Notch signal transduction pathway mediates important cellular functions through direct cell-to-cell contact. Deregulation of Notch activity can lead to an altered cell proliferation and has been linked to many human cancers. Casein kinase 2 (CK2), a ubiquitous kinase, regulates several cellular processes by phosphorylating proteins involved in signal transduction, gene expression, and protein synthesis. In this report we identify NotchICD as a novel target of phosphorylation by CK2. Using mapping and mutational studies, we identified serine 1901, located in the ankyrin domain of Notch, as the target amino acid. Interestingly, phosphorylation of serine 1901 by CK2 appears to generate a second phosphorylation site at threonine 1898. Furthermore, threonine 1898 phosphorylation only occurs when Notch forms a complex with Mastermind and CSL. Phosphorylation of both threonine 1898 and serine 1901 resulted in decreased binding of the Notch-Mastermind-CSL ternary complex to DNA and consequently lower transcriptional activity. These data indicate that the phosphorylation of serine 1901 and threonine 1898 negatively regulates Notch function by dissociating the complex from DNA. This study identifies a new component involved in regulation of NotchICD transcriptional activity, reinforcing the notion that a precise and tight regulation is required for this essential signaling pathway.

Original languageEnglish
Pages (from-to)28844-28857
Number of pages14
JournalJournal of Biological Chemistry
Volume286
Issue number33
DOIs
StatePublished - Aug 19 2011

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Ankyrin Repeat
Phosphorylation
Threonine
Casein Kinase II
Serine
Signal transduction
Signal Transduction
Ankyrins
Deregulation
DNA
Cell proliferation
Gene expression
Proteins
Phosphotransferases
Cell Proliferation
Gene Expression
Amino Acids

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Hierarchical phosphorylation within the ankyrin repeat domain defines a phosphoregulatory loop that regulates notch transcriptional activity. / Ranganathan, Prathibha; Vasquez-Del Carpio, Rodrigo; Kaplan, Fred M.; Wang, Hong; Gupta, Ashu; VanWye, Jeffrey D.; Capobianco, Anthony J.

In: Journal of Biological Chemistry, Vol. 286, No. 33, 19.08.2011, p. 28844-28857.

Research output: Contribution to journalArticle

Ranganathan, Prathibha ; Vasquez-Del Carpio, Rodrigo ; Kaplan, Fred M. ; Wang, Hong ; Gupta, Ashu ; VanWye, Jeffrey D. ; Capobianco, Anthony J. / Hierarchical phosphorylation within the ankyrin repeat domain defines a phosphoregulatory loop that regulates notch transcriptional activity. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 33. pp. 28844-28857.
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