PUBLICATION
Comparing protein stabilities during zebrafish embryogenesis
- Authors
- Becker, T., Bossenz, M., Tursun, B., Schlüter, A., Peters, M.A., Becker, C.G., Ostendorff, H.P., and Bach, I.
- ID
- ZDB-PUB-031229-13
- Date
- 2003
- Source
- Methods in cell science : an official journal of the Society for In Vitro Biology 25(1-2): 85-89 (Review)
- Registered Authors
- Becker, Catherina G., Becker, Thomas
- Keywords
- embryogenesis, mRNA injection, protein stability, ubiquitin ligase, zebrafish
- MeSH Terms
-
- Animals
- Cloning, Molecular
- Embryo, Nonmammalian/metabolism
- Gene Expression Regulation, Developmental/physiology*
- Microinjections
- Protein Denaturation
- RNA Stability/physiology*
- RNA, Messenger/metabolism*
- Repressor Proteins/metabolism*
- Zebrafish/embryology
- Zebrafish/metabolism*
- PubMed
- 14739592 Full text @ Methods Cell Sci.
Citation
Becker, T., Bossenz, M., Tursun, B., Schlüter, A., Peters, M.A., Becker, C.G., Ostendorff, H.P., and Bach, I. (2003) Comparing protein stabilities during zebrafish embryogenesis. Methods in cell science : an official journal of the Society for In Vitro Biology. 25(1-2):85-89.
Abstract
The stabilities of many key proteins are regulated, e.g. via ubiquitination and proteasomal degradation, with important biological consequences. We present a convenient method that allows the analysis and comparison of protein stabilities during embryogenesis using early zebrafish development as a model system. Basically, this method involves ectopic overexpression of epitope-tagged proteins via mRNA injections in one-to-four-cell stage embryos and subsequent protein detection after various time points. Indeed, the protein stability of the ubiquitin ligase RLIM, which is able to autoubiquitinate and target itself for proteasomal degradation, was much shorter when compared to a protein consisting of a Myc epitope-tag and a nuclear localization domain. Thus, this method may be used more widely for the study of developmental protein stability.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping