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

Gen-Der Chen; Chih-Ming Chou; Sheng-Ping L Hwang; Fung-Fang Wang; Yi-Chung Chen; Chin-Chun Hung; Jeou-Yuan Chen; Chang-Jen Huang

doi:10.1016/j.bbrc.2006.03.136

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

Biochem Biophys Res Commun. 2006 May 26;344(1):272-82. doi: 10.1016/j.bbrc.2006.03.136. Epub 2006 Mar 30.

Authors

Gen-Der Chen¹, Chih-Ming Chou, Sheng-Ping L Hwang, Fung-Fang Wang, Yi-Chung Chen, Chin-Chun Hung, Jeou-Yuan Chen, Chang-Jen Huang

Affiliation

¹ Institute of Biochemistry, National Yang-Ming University, Taipei, Taiwan.

PMID: 16616005
DOI: 10.1016/j.bbrc.2006.03.136

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis*
Biological Assay
Cell Nucleus / chemistry
Cell Nucleus / metabolism*
Cytosol / chemistry
Cytosol / metabolism
ELAV Proteins / genetics
ELAV-Like Protein 3
Embryo, Nonmammalian / cytology
Embryo, Nonmammalian / metabolism
Green Fluorescent Proteins / analysis
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Karyopherins / metabolism
Mutation
Neurons / chemistry
Neurons / metabolism
Nuclear Localization Signals / genetics
Nuclear Localization Signals / metabolism
Recombinant Fusion Proteins / analysis
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Transcription, Genetic*
Tumor Suppressor Protein p53 / analysis
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism*
Yolk Sac / chemistry
Yolk Sac / cytology
Yolk Sac / metabolism*
Zebrafish / embryology*
Zebrafish / genetics
Zebrafish / metabolism
Zebrafish Proteins / analysis
Zebrafish Proteins / genetics
Zebrafish Proteins / metabolism
bcl-Associated Death Protein / analysis
bcl-Associated Death Protein / genetics
bcl-Associated Death Protein / metabolism

Substances

ELAV Proteins
ELAV-Like Protein 3
Karyopherins
Nuclear Localization Signals
Recombinant Fusion Proteins
Tumor Suppressor Protein p53
Zebrafish Proteins
badb protein, zebrafish
bcl-Associated Death Protein
elavl3 protein, zebrafish
Green Fluorescent Proteins