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1 Graduate School of Comprehensive Human Sciences, Center for Tsukuba Advanced Research Alliance, and ERATO Environmental Research Project, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba 305-8577, Japan
2 Tohoku University Biomedical Engineering Research Organization, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan
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.
* Correspondence: E-mail: masi{at}tara.tsukuba.ac.jp; nmine{at}tubero.tohoku.ac.jp
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