Kubohara Yuzuru, Arai Akiko, Takahashi Katsunori, Hosaka Kohei, Okamoto Koji
Biosignal Research Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi 371-8512, Japan.
Biochem Biophys Res Commun. 2004 Jul 23;320(2):468-73. doi: 10.1016/j.bbrc.2004.05.189.
We have previously identified a stalk-specific wheat germ agglutinin (WGA)-binding protein, wst34, in the cellular slime mould Dictyostelium discoideum [Biochem. Cell Biol. 68 (1990) 699]. Here, we found another stalk-specific WGA-binding protein, wst25, which was detected with two antisera that recognize wst34. Using the two marker proteins, we then analyzed and compared the pathways of prestalk-to-stalk maturation and prespore-to-stalk conversion in vitro and in vivo. Prestalk cells isolated from normally formed slugs can be converted to stalk cells (designated StI) in vitro with 8-bromo-cAMP (Br-cAMP), whereas prespore cells isolated from slugs can be converted to fully vacuolated stalk cells (designated StII) in vitro with Br-cAMP and DIF-1. During the process of prespore-to-stalk conversion, prespore-specific mRNAs, D19 and 2H3, disappeared rapidly, while prestalk-specific mRNAs, ecmA and ecmB, appeared at 2h of incubation and increased thereafter. Most importantly, however, the StII cells thus formed were biochemically different from the StI cells originated from prestalk cells; that is, StI cells expressed wst34 but not wst25, while StII cells expressed wst25 but not wst34. When prespore cells isolated from slugs were allowed to develop on a substratum, they differentiated into spores and stalk cells and formed fruiting bodies, and the stalk cells formed from prespore cells in vivo expressed wst25 but not wst34. The present results indicate that there are two types of stalk cells, StI (prestalk-origin) and StII (prespore-origin), and that wst34 and wst25 are the specific markers for StI and StII, respectively.
我们之前在细胞黏菌盘基网柄菌中鉴定出一种茎特异性小麦胚凝集素(WGA)结合蛋白wst34[《生物化学与细胞生物学》68(1990)699]。在此,我们发现了另一种茎特异性WGA结合蛋白wst25,它可被两种识别wst34的抗血清检测到。然后,我们使用这两种标记蛋白,在体外和体内分析并比较了前柄细胞到茎细胞的成熟途径以及前孢子细胞到茎细胞的转化途径。从正常形成的蛞蝓中分离出的前柄细胞在体外可通过8-溴环磷酸腺苷(Br-cAMP)转化为茎细胞(称为StI),而从蛞蝓中分离出的前孢子细胞在体外可通过Br-cAMP和DIF-1转化为完全空泡化的茎细胞(称为StII)。在前孢子细胞到茎细胞的转化过程中,前孢子特异性mRNA D19和2H3迅速消失,而前柄特异性mRNA ecmA和ecmB在孵育2小时时出现并随后增加。然而,最重要的是,这样形成的StII细胞在生化性质上与源自前柄细胞的StI细胞不同;也就是说,StI细胞表达wst34但不表达wst25,而StII细胞表达wst25但不表达wst34。当从蛞蝓中分离出的前孢子细胞在基质上发育时,它们分化为孢子和茎细胞并形成子实体,并且在体内由前孢子细胞形成的茎细胞表达wst25但不表达wst34。目前的结果表明存在两种类型的茎细胞,StI(源自前柄)和StII(源自前孢子),并且wst34和wst25分别是StI和StII的特异性标记。