Comparing protein stabilities during zebrafish embryogenesis

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Thomas Becker - , Universität Hamburg (Autor:in)
  • Michael Bossenz - , Universität Hamburg (Autor:in)
  • Baris Tursun - , Universität Hamburg (Autor:in)
  • Anne Schlüter - , Universität Hamburg (Autor:in)
  • Marvin A. Peters - , Universität Hamburg (Autor:in)
  • Catherina G. Becker - , Universität Hamburg (Autor:in)
  • Heather P. Ostendorff - , Universität Hamburg (Autor:in)
  • Ingolf Bach - , Universität Hamburg (Autor:in)

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.

Details

OriginalspracheEnglisch
Seiten (von - bis)85-89
Seitenumfang5
FachzeitschriftMethods in cell science : an official journal of the Society for In Vitro Biology
Jahrgang25
Ausgabenummer1-2
PublikationsstatusVeröffentlicht - 2003
Peer-Review-StatusJa
Extern publiziertJa

Schlagworte

ASJC Scopus Sachgebiete

Schlagwörter

  • Embryogenesis, mRNA injection, Protein stability, Ubiquitin ligase, Zebrafish

Bibliotheksschlagworte