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Identification and differential expression of multiple isoforms of mouse Coiled-coil-DIX1 (Ccd1), a positive

Kensuke Shiomi1, Mizuki Kanemoto, Kazuko Keino-Masu

  • 1Department of Molecular Neurobiology, Institute of Basic Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8577, Japan.

Brain Research. Molecular Brain Research
|April 29, 2005
PubMed

Insights

Mouse Coiled-coil-DIX1 (Ccd1) acts as a positive regulator in the Wnt/beta-catenin signaling pathway. It enhances Wnt pathway activation, particularly in neural development and plasticity.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Cell Signaling

Background:

  • Wnt signaling is crucial for cell growth, differentiation, and neural development.
  • Dishevelled (Dvl) and Axin proteins regulate the canonical Wnt pathway by controlling beta-catenin degradation.
  • Coiled-coil-DIX1 (Ccd1) is a newly identified DIX domain-containing protein involved in Wnt pathway activation.

Purpose of the Study:

  • To isolate and characterize mouse Coiled-coil-DIX1 (Ccd1).
  • To investigate the role of Ccd1 in the Wnt/beta-catenin signaling pathway.
  • To explore the expression patterns and potential functions of Ccd1 in the brain.

Main Methods:

  • Isolation and characterization of mouse Ccd1 and its isoforms.
  • Analysis of mRNA isoforms generated by alternative splicing and promoter usage.
  • Co-expression studies in HeLa cells to assess Ccd1's effect on TCF-dependent reporter transcription.
  • Inhibition assays using Axin or dominant-negative Ccd1 to study pathway regulation.
  • Immunohistochemical analysis to determine Ccd1 expression and localization in the brain.

Main Results:

  • Fourteen putative mRNA isoforms of mouse Ccd1 were identified, with distinct tissue expression patterns.
  • Ccd1 proteins were classified into three subtypes, including a novel Ccd1A isoform with a calponin homology domain.
  • Ccd1 alone showed minimal Wnt pathway activation, but potentiated Dvl-induced activation in a dose-dependent manner.
  • Axin or dominant-negative Ccd1 inhibited Ccd1- and Wnt3a-dependent Wnt pathway activation.
  • Ccd1 is highly expressed and co-localized with Wnt signaling molecules in the embryonic and adult brain.

Conclusions:

  • Mouse Ccd1 functions as a positive regulator of the Wnt/beta-catenin pathway.
  • Ccd1 isoforms exhibit diverse expression profiles, suggesting specialized roles.
  • The novel Ccd1A isoform may interact with the actin cytoskeleton.
  • Ccd1 plays a significant role in Wnt-mediated neuronal development, plasticity, and remodeling.

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