Back to Search
Start Over
Cloning of zebrafish BAD, a BH3-only proapoptotic protein, whose overexpression leads to apoptosis in COS-1 cells and zebrafish embryos
- Source :
- Biochemical and biophysical research communications. 304(4)
- Publication Year :
- 2003
-
Abstract
- The BH3-only proapoptotic protein, BAD, was cloned from zebrafish embryos and its properties were characterized. Zebrafish BAD (zBAD) is a protein with 147 amino acids that contains a BH3 domain and a putative 14-3-3 binding site with the sequence of RPRSRS84AP, corresponding to S136 in mouse BAD (mBAD). zBAD shares 34%, 28%, and 29% amino acid sequence identity to the human, mouse, and rat BAD, respectively. RT-PCR analysis revealed that the expression of zBAD gene is found in various parts of zebrafish tissues. The treatment with the z-VAD fmk, a broad-range caspase inhibitor, in COS-1 cells significantly increased the expression of zebrafish BAD fusion proteins (GFP-zBAD and HA-zBAD), indicating that zebrafish BAD fusion proteins may be cleaved by caspase(s). zBAD was shown to induce apoptosis when it was overexpressed in COS-1 cells. In addition, zBAD was also expressed in muscle cells under the muscle-specific promoter from zebrafish α-actin gene. Abnormality in the skeletal muscles and the loss of green fluorescence signal in the same region were observed. Taken together, our results indicate that zBAD could induce apoptosis in vitro and in vivo and may have biological implications in apoptosis during zebrafish development.
- Subjects :
- Recombinant Fusion Proteins
Molecular Sequence Data
Biophysics
Apoptosis
Cysteine Proteinase Inhibitors
Biochemistry
Amino Acid Chloromethyl Ketones
Genes, Reporter
Animals
Humans
Amino Acid Sequence
Cloning, Molecular
Molecular Biology
Gene
Peptide sequence
Zebrafish
Caspase
chemistry.chemical_classification
biology
Cell Biology
Zebrafish Proteins
biology.organism_classification
Molecular biology
Fusion protein
In vitro
Actins
Amino acid
chemistry
COS Cells
biology.protein
bcl-Associated Death Protein
Carrier Proteins
Sequence Alignment
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 304
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- Biochemical and biophysical research communications
- Accession number :
- edsair.doi.dedup.....639ffcf333c950b4320e367515a95afb