Rapid turnover of GATA-2 via ubiquitin-proteasome protein degradation pathway

Genes Cells. 2005 Jul;10(7):693-704. doi: 10.1111/j.1365-2443.2005.00864.x.

Abstract

Transcription factor GATA-2 is expressed in a number of tissues, including hematopoietic stem and progenitor cells, and is crucial for the proliferation and survival of hematopoietic cells. To further characterize the function of GATA-2, we examined the cellular turnover mechanism of GATA-2. In P815 cells, the half-life of endogenous GATA-2 was found to be as short as 30 min after cycloheximide treatment. This short half-life was reproducible in other hematopoietic and neuroblastoma cell lines with moderate variation. We also found that ultraviolet (UV)-C irradiation markedly represses the GATA-2 protein level by facilitating the degradation process. Since treatment of the cells with the proteasome inhibitor MG132 or clasto-Lactacystin substantially abrogated the effects of cycloheximide and UV-C irradiation and increased the expression level of both endogenous and transfected GATA-2, the degradation of GATA-2 seems to occur through the proteasome pathway. Structure-function analyses with the GAL4-DNA binding domain (GBD)-GATA-2 fusion protein and GATA-2 deletion mutants suggested that the protein degradation regulatory elements of GATA-2 reside in three regions, two of which overlap with the transactivation domain. We also detected poly ubiquitinated forms of GATA-2. Taken together, these results demonstrate that GATA-2 is turned over rapidly through the ubiquitin-proteasome pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / analogs & derivatives
  • Acetylcysteine / pharmacology
  • Animals
  • Cells, Cultured / drug effects
  • Cells, Cultured / radiation effects
  • Cycloheximide / pharmacology
  • Cysteine Proteinase Inhibitors / pharmacology
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • GATA2 Transcription Factor
  • Gene Deletion
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / radiation effects
  • Half-Life
  • Humans
  • Leukemia, Experimental / metabolism*
  • Leukemia, Experimental / pathology
  • Leupeptins / pharmacology
  • Mice
  • Mutation
  • Neuroblastoma / metabolism*
  • Neuroblastoma / pathology
  • Proteasome Endopeptidase Complex / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcriptional Activation
  • Ubiquitins / metabolism*
  • Ultraviolet Rays

Substances

  • Cysteine Proteinase Inhibitors
  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • GATA2 Transcription Factor
  • GATA2 protein, human
  • Gata2 protein, mouse
  • Leupeptins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Ubiquitins
  • lactacystin
  • Cycloheximide
  • Proteasome Endopeptidase Complex
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde
  • Acetylcysteine