Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, 2-1 Seiryo-cho, Aoba-ku, Sendai 980-8575, Japan.
J Biol Chem. 2010 Jan 1;285(1):773-83. doi: 10.1074/jbc.M109.030726. Epub 2009 Oct 23.
GATA1 is essential for the differentiation of erythroid cells and megakaryocytes. The Gata1 gene is composed of multiple untranslated first exons and five common coding exons. The erythroid first exon (IE exon) is important for Gata1 gene expression in hematopoietic lineages. Because previous IE exon knockdown analyses resulted in embryonic lethality, less is understood about the contribution of the IE exon to adult hematopoiesis. Here, we achieved specific deletion of the floxed IE exon in adulthood using an inducible Cre expression system. In this conditional knock-out mouse line, the Gata1 mRNA level was significantly down-regulated in the megakaryocyte lineage, resulting in thrombocytopenia with a marked proliferation of megakaryocytes. By contrast, in the erythroid lineage, Gata1 mRNA was expressed abundantly utilizing alternative first exons. Especially, the IEb/c and newly identified IEd exons were transcribed at a level comparable with that of the IE exon in control mice. Surprisingly, in the IE-null mouse, these transcripts failed to produce full-length GATA1 protein, but instead yielded GATA1 lacking the N-terminal domain inefficiently. With low level expression of the short form of GATA1, IE-null mice showed severe anemia with skewed erythroid maturation. Notably, the hematological phenotypes of adult IE-null mice substantially differ from those observed in mice harboring conditional ablation of the entire Gata1 gene. The present study demonstrates that the IE exon is instrumental to adult erythropoiesis by regulating the proper level of transcription and selecting the correct transcription start site of the Gata1 gene.
GATA1 对于红细胞和巨核细胞的分化是必不可少的。Gata1 基因由多个未翻译的第一外显子和五个常见的编码外显子组成。红细胞第一外显子(IE 外显子)对于造血谱系中 Gata1 基因的表达很重要。由于之前的 IE 外显子敲低分析导致胚胎致死,因此对 IE 外显子对成体造血的贡献了解较少。在这里,我们使用诱导型 Cre 表达系统在成年期实现了 floxed IE 外显子的特异性缺失。在这种条件性敲除小鼠品系中,Gata1 mRNA 水平在巨核细胞谱系中显著下调,导致血小板减少症,巨核细胞大量增殖。相比之下,在红细胞谱系中,Gata1 mRNA 大量利用替代的第一外显子表达。特别是,IEb/c 和新鉴定的 IEd 外显子的转录水平与对照小鼠中的 IE 外显子相当。令人惊讶的是,在 IE 缺失的小鼠中,这些转录物未能产生全长的 GATA1 蛋白,而是产生缺乏 N 端结构域的 GATA1 蛋白效率低下。由于短形式 GATA1 的低水平表达,IE 缺失的小鼠表现出严重的贫血和红细胞成熟偏斜。值得注意的是,成年 IE 缺失小鼠的血液学表型与那些携带 Gata1 基因完全缺失的小鼠观察到的表型有很大的不同。本研究表明,IE 外显子通过调节 Gata1 基因的适当转录水平和选择正确的转录起始位点,对成体红细胞生成至关重要。