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

机译：通过泛素-蛋白酶体蛋白降解途径快速转换GATA-2。

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摘要

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.
机译：转录因子GATA-2在许多组织中表达，包括造血干细胞和祖细胞，并且对造血细胞的增殖和存活至关重要。为了进一步表征GATA-2的功能，我们研究了GATA-2的细胞更新机制。在P815细胞中，发现内源性GATA-2的半衰期短于环己酰亚胺处理后的30分钟。这种短的半衰期在其他造血和成神经细胞瘤细胞系中具有中等变异性，可再现。我们还发现，紫外线（UV）-C辐射可通过促进降解过程显着抑制GATA-2蛋白水平。由于用蛋白酶体抑制剂MG132或clasto-Lactacystin处理细胞可基本消除环己酰亚胺和UV-C辐射的影响，并增加内源性和转染的GATA-2的表达水平，因此似乎通过GATA-2降解发生了蛋白酶体途径。 GAL4-DNA结合域（GBD）-GATA-2融合蛋白和GATA-2缺失突变体的结构功能分析表明，GATA-2的蛋白质降解调控元件位于三个区域，其中两个与反式激活域重叠。我们还检测了GATA-2的多泛素化形式。综上所述，这些结果表明，GATA-2通过泛素-蛋白酶体途径迅速被翻转。

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