Richardson D L, Hong C B, Loomis W F
Department of Biology, University of California, San Diego, La Jolla 92093-0322.
Dev Biol. 1991 Apr;144(2):269-80. doi: 10.1016/0012-1606(91)90421-x.
During culmination of Dictyostelium fruiting bodies, prespore and prestalk cells undergo terminal differentiation to form spores and a cellular stalk. A genomic fragment was isolated by random cloning that hybridizes to a 1.4-kb mRNA present during culmination. Cell type separations at culmination showed that the mRNA is present in prespore cells and spores, but not in prestalk or stalk cells. After genomic mapping, an additional 3 kb of DNA surrounding the original 1-kb fragment was cloned. The gene was sequenced and named Dd31 after the size of the predicted protein product in kilodaltons. Accumulation of Dd31 mRNA occurs immediately prior to sporulation. Addition of 20 mM 8-Br-cAMP to cells dissociated from Mexican hat stage culminants induced sporulation and the accumulation of Dd31 mRNA, while 20 mM cAMP did not. Dd31 mRNA does not accumulate in the homeotic mutant stalky in which prespore cells are converted to stalk cells rather than spores. Characterization of Dd31 extends the known temporal dependent sequence of molecular differentiations to sporulation.
在盘基网柄菌子实体发育成熟过程中,前孢子细胞和前柄细胞经历终末分化形成孢子和细胞性柄。通过随机克隆分离出一个基因组片段,它与发育成熟过程中出现的一种1.4 kb的mRNA杂交。发育成熟时的细胞类型分离显示,该mRNA存在于前孢子细胞和孢子中,但不存在于前柄细胞或柄细胞中。经过基因组定位后,围绕最初的1 kb片段又克隆了另外3 kb的DNA。对该基因进行测序后,根据预测蛋白质产物的千道尔顿大小将其命名为Dd31。Dd31 mRNA的积累发生在孢子形成之前不久。向从墨西哥帽阶段发育成熟体解离的细胞中添加20 mM 8-溴环腺苷酸可诱导孢子形成和Dd31 mRNA的积累,而20 mM环腺苷酸则无此作用。Dd31 mRNA在同源异型突变体“stalky”中不积累,在该突变体中,前孢子细胞转化为柄细胞而非孢子。对Dd31的表征将已知的分子分化时间依赖性序列扩展到了孢子形成过程。