Hoe K L, Won M S, Chung K S, Park S K, Kim D U, Jang Y J, Yoo O J, Yoo H S
Cell Cycle, Signal Transduction Research Unit, Korea Research Institute of Bioscience, Biotechnology, Taejon 305-606, South Korea.
Gene. 1998 Jul 30;215(2):319-28. doi: 10.1016/s0378-1119(98)00301-1.
As a first step to elucidate the functions of Schizosaccharomyces pombe (S. pombe) GATA factors, we have isolated the gaf1+ gene (GATA-factor like gene) in S. pombe. The predicted amino acid (aa) sequence of Gaf1 reveals a single zinc finger domain typical of fungal GATA factors, and the zinc finger exhibits 60% aa identity to that of human GATA-1. The open reading frame of Gaf1 predicts a protein of Mr 32 kDa consisting of 290 intronless amino acids. Disruption of this gene has no effect on cell viability and growth rate. The GST-Gaf1 fusion protein binds specifically to GATA motifs of its own promoter as well as DAL7 UAS, a canonical GATA motif of Saccharomyces cerevisiae (S. cerevisiae) The specific DNA-binding activity resides within the N-terminal half of Gaf1 (Gaf1N; aa 1-120) containing the zinc finger, whereas the C-terminal half (Gaf1C; aa 121-290) contains transactivation sequences that induce the expression of the lacZ reporter when fused to the GAL4 DNA binding domain. These results demonstrate that Gaf1 may function as a transcriptional activator consisting of DNA-binding and transactivation domains.
作为阐明粟酒裂殖酵母（裂殖酵母）GATA因子功能的第一步，我们已在裂殖酵母中分离出gaf1 +基因（类GATA因子基因）。Gaf1预测的氨基酸序列显示出真菌GATA因子典型的单个锌指结构域，并且该锌指与人GATA-1的锌指具有60％的氨基酸同一性。Gaf1的开放阅读框预测一个32 kDa的蛋白质，由290个无内含子氨基酸组成。该基因的破坏对细胞活力和生长速率没有影响。GST-Gaf1融合蛋白特异性结合其自身启动子的GATA基序以及酿酒酵母（酿酒酵母）的典型GATA基序DAL7 UAS。特异性DNA结合活性存在于包含锌指的Gaf1的N端一半（Gaf1N；氨基酸1-120）内，而C端一半（Gaf1C；氨基酸121-290）包含当与GAL4 DNA结合结构域融合时诱导lacZ报告基因表达的反式激活序列。这些结果表明，Gaf1可能作为由DNA结合结构域和反式激活结构域组成的转录激活因子发挥作用。
