Richardson D L, Loomis W F, Kimmel A R
Laboratory of Cellular and Developmental Biology, NIDDK, NIH, Bethesda, MD 20892, USA.
Development. 1994 Oct;120(10):2891-900. doi: 10.1242/dev.120.10.2891.
spiA, a marker for sporulation, is expressed during the culmination stage of Dictyostelium development, when the mass of prespore cells has moved partly up the newly formed stalk. Strains containing a full-length spiA promoter/lacZ fusion were stained for beta-galactosidase activity at intervals during development. The results indicate that expression of spiA initiates in prespore cells at the prestalk/prespore boundary (near the apex) and extends downward into the prespore mass as culmination continues. A spatial gradient of staining expands from the top of the prespore mass and intensifies until the front of activation reaches the bottom, whereupon the entire region stains darkly. The spiA promoter can be deleted to within 301 bp of the transcriptional start site with no effect on the relative strength, timing or spatial localization of expression. Further 5' deletions from -301 to -175 reduce promoter strength incrementally, although timing and spatial expression are not affected. Deletions to -159 and beyond result in inactive promoters. Treatment of early developmental structures with 8-Br-cAMP in situ activates the intracellular cAMP-dependent protein kinase (PKA) and precociously induces spiA expression and sporulation. The absence of an apparent gradient of staining in these structures suggest that PKA is equivalently activatable throughout the prespore region and that all prespore cells are competent to express spiA. Thus, we postulate that the pattern of expression of spiA reveals the progression of an inductive signal for sporulation and suggest that this signal may originate from the prestalk cells at the apex.
spiA是一种孢子形成的标志物,在盘基网柄菌发育的 culmination 阶段表达,此时前孢子细胞团已部分向上移动到新形成的柄上。在发育过程中,对含有全长spiA启动子/lacZ融合基因的菌株进行β-半乳糖苷酶活性染色。结果表明,spiA的表达在前柄/前孢子边界(靠近顶端)的前孢子细胞中开始,并随着 culmination 的继续向下延伸到前孢子团中。染色的空间梯度从前孢子团的顶部扩展并增强,直到激活前沿到达底部,此时整个区域染色加深。spiA启动子可以缺失到转录起始位点的301 bp以内,而对表达的相对强度、时间或空间定位没有影响。从-301到-175的进一步5'缺失会逐渐降低启动子强度,尽管时间和空间表达不受影响。缺失到-159及更远会导致启动子失活。用8-Br-cAMP原位处理早期发育结构可激活细胞内cAMP依赖性蛋白激酶(PKA),并早熟诱导spiA表达和孢子形成。这些结构中没有明显的染色梯度表明PKA在整个前孢子区域都可等效激活,并且所有前孢子细胞都有能力表达spiA。因此,我们推测spiA的表达模式揭示了孢子形成诱导信号的进展,并表明该信号可能起源于顶端的前柄细胞。