PUBLICATION

Requirement of nuclear localization and transcriptional activity of p53 for its targeting to the yolk syncytial layer (YSL) nuclei in zebrafish embryo and its use for apoptosis assay

Authors
Chen, G.D., Chou, C.M., Hwang, S.P., Wang, F.F., Chen, Y.C., Hung, C.C., Chen, J.Y., and Huang, C.J.
ID
ZDB-PUB-060419-7
Date
2006
Source
Biochemical and Biophysical Research Communications   344(1): 272-282 (Journal)
Registered Authors
Huang, Chang-Jen, Hwang, Sheng-Ping L., Wang, Chunping
Keywords
p53, Zebrafish, Yolk syncytial layer, Apoptosis, Nuclear localization, Neuron-specific promoter
MeSH Terms
  • Nuclear Localization Signals/genetics
  • Nuclear Localization Signals/metabolism
  • Karyopherins/metabolism
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/metabolism
  • Transcription, Genetic*
  • Neurons/chemistry
  • Neurons/metabolism
  • Yolk Sac/chemistry
  • Yolk Sac/cytology
  • Yolk Sac/metabolism*
  • Biological Assay
  • bcl-Associated Death Protein/analysis
  • bcl-Associated Death Protein/genetics
  • bcl-Associated Death Protein/metabolism
  • ELAV-Like Protein 3
  • Animals
  • Zebrafish Proteins/analysis
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism
  • ELAV Proteins/genetics
  • Zebrafish/embryology*
  • Zebrafish/genetics
  • Zebrafish/metabolism
  • Cell Nucleus/chemistry
  • Cell Nucleus/metabolism*
  • Mutation
  • Tumor Suppressor Protein p53/analysis
  • Tumor Suppressor Protein p53/genetics
  • Tumor Suppressor Protein p53/metabolism*
  • Green Fluorescent Proteins/analysis
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism
  • Recombinant Fusion Proteins/analysis
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/metabolism
  • Apoptosis*
  • Cytosol/chemistry
  • Cytosol/metabolism
PubMed
16616005 Full text @ Biochem. Biophys. Res. Commun.
Abstract
We expressed zebrafish p53 protein fused to GFP by a neuron-specific HuC promoter in zebrafish embryos. Instead of displaying neuronal expression patterns, p53-GFP was targeted to zebrafish YSL nuclei. This YSL targeting is p53 sequence-specific because GFP fusion proteins of p63 and p73 displayed neuronal-specific patterns. To dissect the underlying mechanisms, various constructs encoding a series of p53 mutant proteins under the control of different promoters were generated. Our results showed that expression of p53, in early zebrafish embryo, is preferentially targeted to the nuclei of YSL, which is mediated by importin. Similarly, this targeting is abrogated when p53 nuclear localization signal is disrupted. In addition, the transcriptional activity of p53 is required for this targeting. We further showed that fusion of pro-apoptotic BAD protein to p53-GFP led to apoptosis of YSL cells, and subsequent imperfect microtubule formation and abnormal blastomere movements.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping