NARF, an nemo-like kinase (NLK)-associated ring finger protein regulates the ubiquitylation and degradation of T cell factor/lymphoid enhancer factor (TCF/LEF)

J Biol Chem. 2006 Jul 28;281(30):20749-20760. doi: 10.1074/jbc.M602089200. Epub 2006 May 19.

Abstract

beta-Catenin is a key player in the Wnt signaling pathway, and interacts with cofactor T cell factor/lymphoid enhancer factor (TCF/LEF) to generate a transcription activator complex that activates Wnt-induced genes. We previously reported that Nemo-like kinase (NLK) negatively regulates Wnt signaling via phosphorylation of TCF/LEF. To further evaluate the physiological roles of NLK, we performed yeast two-hybrid screening to identify NLK-interacting proteins. From this screen, we isolated a novel RING finger protein that we term NARF (NLK associated RING finger protein). Here, we show that NARF induces the ubiquitylation of TCF/LEF in vitro and in vivo, and functions as an E3 ubiquitin-ligase that specifically cooperates with the E2 conjugating enzyme E2-25K. We found that NLK augmented NARF binding and ubiquitylation of TCF/LEF, and this required NLK kinase activity. The ubiquitylated TCF/LEF was subsequently degraded by the proteasome. Furthermore, NARF inhibited formation of the secondary axis induced by the ectopic expression of beta-catenin in Xenopus embryos. Collectively, our findings raise the possibility that NARF functions as a novel ubiquitin-ligase to suppress the Wnt-beta-catenin signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / biosynthesis*
  • Oocytes / metabolism
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • TCF Transcription Factors / metabolism*
  • Two-Hybrid System Techniques
  • Ubiquitin / chemistry*
  • Ubiquitin / metabolism
  • Xenopus
  • beta Catenin / metabolism

Substances

  • NARF protein, human
  • Nuclear Proteins
  • TCF Transcription Factors
  • Ubiquitin
  • beta Catenin