PUBLICATION
Low-molecular-weight vitellogenin 1-like proteins are components of a UV-damaged-DNA binding activity highly expressed in zebrafish (Danio rerio) embryos
- Authors
- Lai, Y.S., Chiue, L.F., and Hsu, T.
- ID
- ZDB-PUB-060130-4
- Date
- 2006
- Source
- Journal of experimental zoology. Part A, Ecological genetics and physiology 305(3): 215-224 (Journal)
- Registered Authors
- Hsu, Todd
- Keywords
- none
- MeSH Terms
-
- Animals
- Blotting, Western
- Chelating Agents/pharmacology
- DNA/metabolism*
- DNA/radiation effects
- DNA Damage/physiology*
- DNA-Binding Proteins/metabolism*
- Electrophoretic Mobility Shift Assay
- Female
- Male
- Peptide Mapping
- Phenanthrolines/pharmacology
- Pyrimidine Dimers/metabolism*
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Vitellogenins/genetics
- Vitellogenins/metabolism*
- Zebrafish/embryology*
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 16432884 Full text @ J. Exp. Zool. Part A Ecol. Genet. Physiol.
Citation
Lai, Y.S., Chiue, L.F., and Hsu, T. (2006) Low-molecular-weight vitellogenin 1-like proteins are components of a UV-damaged-DNA binding activity highly expressed in zebrafish (Danio rerio) embryos. Journal of experimental zoology. Part A, Ecological genetics and physiology. 305(3):215-224.
Abstract
A strong UV-damaged-DNA binding activity had been detected in the extracts of zebrafish embryos at 12 hr after fertilization by gel shift assay (Hsu et al. 2002. Fish Physiol Biochem 25:41-51). We attempted to study the components of this binding activity and their importance in DNA damage recognition. Among the proteins extracted from gel retardation complexes, a 30- and a 35-kDa polypeptide binding preferentially to 6-4photoproducts (6-4PPs) generated by UV irradiation were identified by peptide mass fingerprinting (PMF) as homologs of zebrafish vitellogenin I (zfVg1), a 150-kDa metalloprotein known as the precursor of yolk proteins in embryos. zfVg1-like polypeptides ranging from 25 to 105 kDa were detected in 12- and 96-hr-old zebrafish extracts by immunoblot analysis. Immunoblot analysis of affinity-captured proteins confirmed the preferential binding of the 30-35-kDa polypeptides to the 6-4PP probe, while 96-hr-old larval extracts containing very low levels of these two factors failed to recognize 6-4PPs. The presence of zfVg1-like factors was important in maintaining the embryonic UV-binding activity, as inclusion of a monoclonal anti-zfVg1 antibody in reaction mixtures caused a concentration-dependent reduction in 6-4PP-specific binding. In contrast, DNA damage recognition was not disturbed at all by an anti-HSP 70 antibody. The formation of 6-4PP-binding complexes was abolished after the addition of the metal chelating agent 1,10-phenanthroline (OP) to zebrafish extracts and the loss of UV-binding capacity correlated with the disappearance of the 35-kDa factor in OP-treated extracts. Our results demonstrated the ability of low-molecular-weight zfVg1-like proteins in zebrafish embryos to bind UV-damaged DNA and the expression of this embryonic UV-binding activity was metal dependent. Whether zfVg1-like UV-binding proteins are involved in repairing damaged DNA in embryos or in processing helical structures similar to UV-distorted DNA needs further investigation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping