Louis S A, Spiegelman G B, Weeks G
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
Mol Biol Cell. 1997 Feb;8(2):303-12. doi: 10.1091/mbc.8.2.303.
It has been previously demonstrated that the expression of an activated rasD gene in wild-type Dictyostelium cells results in formation of aggregates with multitips, instead of the normal single tips, and a block in further development. In an attempt to better understand the role of activated RasD development, we examined cell-type-specific gene expression in a strain stably expressing high levels of RasD[G12T]. We found that the expression of prestalk cell-specific genes ecmA and tagB was markedly enhanced, whereas the expression of the prespore cell-specific gene cotC was reduced to very low levels. When the fate of cells in the multitipped aggregate was monitored with an ecmA/lacZ fusion, it appeared that most of the cells eventually adopted prestalk gene expression characteristics. When mixtures of the [G12T]rasD cells and Ax3 cells were induced to differentiate, chimeric pseudoplasmodia were not formed. Thus, although the [G12T]rasD transformant had a marked propensity to form prestalk cells, it could not supply the prestalk cell population when mixed with wild-type cells. Both stalk and spore cell formation occurred in low cell density monolayers of the [G12T]rasD strain, suggesting that at least part of the inhibition of stalk and spore formation during multicellular development involved inhibitory cell interactions within the cell mass. Models for the possible role of rasD in development are discussed.
先前已经证明,在野生型盘基网柄菌细胞中激活的rasD基因的表达会导致形成具有多个尖端的聚集体,而不是正常的单个尖端,并且会阻碍进一步发育。为了更好地理解激活的RasD在发育中的作用,我们检测了稳定表达高水平RasD[G12T]的菌株中细胞类型特异性基因的表达。我们发现,前柄细胞特异性基因ecmA和tagB的表达明显增强,而前孢子细胞特异性基因cotC的表达降低到非常低的水平。当用ecmA/lacZ融合蛋白监测多尖端聚集体中细胞的命运时,似乎大多数细胞最终都具有前柄基因表达特征。当诱导[G12T]rasD细胞和Ax3细胞的混合物分化时,没有形成嵌合伪原质团。因此,尽管[G12T]rasD转化体具有形成前柄细胞的明显倾向,但当与野生型细胞混合时,它不能提供前柄细胞群体。在[G12T]rasD菌株的低细胞密度单层中同时发生了柄细胞和孢子细胞的形成,这表明在多细胞发育过程中对柄细胞和孢子细胞形成的抑制至少部分涉及细胞团内的抑制性细胞相互作用。本文讨论了rasD在发育中可能作用的模型。