PUBLICATION

Sinup, a novel Siaz-interacting nuclear protein, modulates neural plate formation in the zebrafish embryos

Authors
Ro, H., Won, M., Lee, S.U., Kim, K.E., Huh, T.L., Kim, C.H., and Rhee, M.
ID
ZDB-PUB-050603-9
Date
2005
Source
Biochemical and Biophysical Research Communications   332(4): 993-1003 (Journal)
Registered Authors
Huh, Tae-Lin, Kim, Cheol-Hee
Keywords
Siaz; Sinup; Yeast two-hybrid assay; Neural plate; Zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Carrier Proteins/metabolism
  • Carrier Proteins/physiology*
  • Cell Nucleus/metabolism
  • Cytoskeletal Proteins/biosynthesis
  • DNA, Complementary/metabolism
  • Ectoderm/metabolism
  • Gene Expression Regulation, Developmental*
  • Gene Library
  • Glutathione Transferase/metabolism
  • Green Fluorescent Proteins/metabolism
  • Homeodomain Proteins/biosynthesis
  • Humans
  • In Situ Hybridization
  • Molecular Sequence Data
  • Neural Crest/embryology*
  • Nuclear Proteins/chemistry
  • Nuclear Proteins/metabolism*
  • Oligonucleotides/chemistry
  • Otx Transcription Factors
  • Plasmids/metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA/chemistry
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Time Factors
  • Transfection
  • Two-Hybrid System Techniques
  • Ubiquitin-Protein Ligases
  • Wnt Proteins
  • Zebrafish
  • Zebrafish Proteins/biosynthesis
  • Zebrafish Proteins/chemistry
  • Zebrafish Proteins/metabolism*
  • Zebrafish Proteins/physiology*
PubMed
15922293 Full text @ Biochem. Biophys. Res. Commun.
Abstract
Siah, the vertebrate homologue of the Drosophila seven in absentia (sina) gene, is well conserved from Drosophila to mammal and involved in ubiquitination and proteasome-dependent degradation of various target proteins. To identify cellular proteins interacting with Siah, we screened a zebrafish cDNA library with zebrafish Siah (Siaz) as bait in a yeast two-hybrid assay. We identified a cDNA encoding a novel protein composed of 145 amino acids and termed it as Sinup (Siaz-interacting-nuclear-protein). Sinup is a novel nuclear protein that binds to the highly conserved C-terminal protein-interacting domain of Siaz both in vivo and in vitro. During development, sinup transcripts are abundant from the one-cell stage to the early blastula and then markedly diminished, suggesting sinup largely exists as maternal transcripts. sinup overexpression induced lateral expansion of the neural plate and in consequence caused ectopic expression of otx-2 and hoxb1b during the late gastrula stage. In addition, the lateral/paraxial expression of wnt8 at the onset of gastrulation is suppressed by the forced expression of sinup while the expression levels of various dorso-ventral markers are unaffected. In contrast, interfering with sinup functions using sinup morpholino oligonucleotides gradually diminished the anterior neuroectoderm from the posterior region, and resulted in compete loss of hindbrain at the 3-somites stage. Our report suggests that sinup expression should be tightly regulated during early embryonic development for the proper neural plate formation.
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