Shaulsky G, Escalante R, Loomis W F
Department of Biology, University of California at San Diego, La Jolla 92093, USA.
Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15260-5. doi: 10.1073/pnas.93.26.15260.
We have found conditions for saturation mutagenesis by restriction enzyme mediated integration that result in plasmid tagging of disrupted genes. Using this method we selected for mutations in genes that act at checkpoints downstream of the intercellular signalling system that controls encapsulation in Dictyostelium discoideum. One of these genes, mkcA, is a member of the mitogen-activating protein kinase cascade family while the other, regA is a novel bipartite gene homologous to response regulators in one part and to cyclic nucleotide phosphodiesterases in the other part. Disruption of either of these genes results in partial suppression of the block to spore formation resulting from the loss of the prestalk genes, tagB and tagC. The products of the tag genes have conserved domains of serine protease attached to ATP-driven transporters, suggesting that they process and export peptide signals. Together, these genes outline an intercellular communication system that coordinates organismal shape with cellular differentiation during development.
我们已经找到了通过限制酶介导的整合进行饱和诱变的条件，该条件可导致对破坏基因进行质粒标记。利用这种方法，我们筛选了在盘基网柄菌中控制包囊化的细胞间信号系统下游检查点起作用的基因突变。其中一个基因mkcA是丝裂原活化蛋白激酶级联家族的成员，而另一个基因regA是一个新的二分体基因，一部分与应答调节因子同源，另一部分与环核苷酸磷酸二酯酶同源。破坏这两个基因中的任何一个都会部分抑制因前柄基因tagB和tagC缺失而导致的孢子形成障碍。tag基因的产物具有与ATP驱动转运蛋白相连的丝氨酸蛋白酶保守结构域，这表明它们加工并输出肽信号。这些基因共同勾勒出一个细胞间通讯系统，该系统在发育过程中协调生物体形态与细胞分化。
