Ogasawara Shun, Shimada Nao, Kawata Takefumi
Department of Biology, Faculty of Science, Toho University, 2-2-1 Miyama, Funabashi, Chiba 274-8510, Japan.
Dev Growth Differ. 2009 Feb;51(2):109-22. doi: 10.1111/j.1440-169X.2009.01086.x.
Expansins are proteins involved in plant morphogenesis, exerting their effects on cellulose to extend cell walls. Dictyostelium is an organism that possesses expansin-like molecules, but their functions are not known. In this study, we analyzed the expL7 (expansin-like 7) gene, which has been identified as a putative target of Dd-STATa, a Dictyostelium homolog of the metazoan signal transducer and activator of transcription (STAT) proteins. Promoter fragments of the expL7 were fused to a lacZ reporter and the expression patterns determined. As expected from the behavior of the endogenous expL7 gene, the expL7/lacZ fusion gene was downregulated in Dd-STATa null slugs. In the parental strain, the expL7 promoter was activated in the anterior tip region. Mutational analysis of the promoter identified a sequence that was necessary for expression in tip cells. In addition, an activator sequence for pstAB cells was identified. These sequences act in combination with the repressor region to prevent ectopic expL7 expression in the prespore and prestalk regions of the slug and culminant. Although the expL7 null mutant showed no phenotypic change, the expL7 overexpressor showed aberrant stalk formation. These results indicate that the expansin-like molecule is important for morphogenesis in Dictyostelium.
扩展蛋白是参与植物形态发生的蛋白质,它们通过作用于纤维素来伸展细胞壁。盘基网柄菌是一种拥有类扩展蛋白分子的生物,但其功能尚不清楚。在本研究中,我们分析了expL7(类扩展蛋白7)基因,该基因已被确定为盘基网柄菌中Dd-STATa的一个假定靶点,Dd-STATa是后生动物信号转导和转录激活因子(STAT)蛋白的盘基网柄菌同源物。将expL7的启动子片段与lacZ报告基因融合,并确定其表达模式。正如内源性expL7基因的行为所预期的那样,expL7/lacZ融合基因在Dd-STATa缺失的蛞蝓体中表达下调。在亲本菌株中,expL7启动子在前尖端区域被激活。对启动子的突变分析确定了一个在尖端细胞中表达所必需的序列。此外,还鉴定了一个pstAB细胞的激活序列。这些序列与抑制区域共同作用,以防止expL7在蛞蝓体和聚集物的前孢子和前柄区域异位表达。尽管expL7基因敲除突变体没有表现出表型变化,但expL7过表达体表现出异常的柄形成。这些结果表明,类扩展蛋白分子对盘基网柄菌的形态发生很重要。