Moraleda-Muñoz Aurelio, Carrero-Lérida Juana, Pérez Juana, Muñoz-Dorado José
Departamento de Microbiología, Facultad de Ciencias, Universidad de Granada, E-18071 Granada, Spain.
J Bacteriol. 2003 Feb;185(4):1376-83. doi: 10.1128/JB.185.4.1376-1383.2003.
We have cloned a two-component regulatory system (phoR2-phoP2) of Myxococcus xanthus while searching for genes that encode proteins with phosphatase activity, where phoR2 encodes the histidine kinase and phoP2 encodes the response regulator. A second system, phoR3-phoP3, was identified and isolated by using phoP2 as a probe. These two systems are quite similar, sharing identities along the full-length proteins of 52% on the histidine kinases and 64% on the response regulators. The predicted structures of both kinases suggest that they are anchored to the membrane, with the sensor domains being located in the periplasmic space and the kinase domains in the cytoplasm. The response regulators (PhoP2 and PhoP3) exhibit a helix-loop-helix motif typical of DNA-binding proteins in the effector domains located in the C-terminal region. Studies on two single-deletion mutants and one double-deletion mutant have revealed that these systems are involved in development. Mutant fruiting bodies are not well packed, originating loose and flat aggregates where some myxospores do not reshape properly, and they remain as elongated cells. These systems are also involved in the expression of Mg-independent acid and neutral phosphatases, which are expressed during development. The neutral phosphatase gene is especially dependent on PhoP3. Neither PhoP2 nor PhoP3 regulates the expression of alkaline phosphatases and the pph1 gene.
在寻找编码具有磷酸酶活性蛋白质的基因时,我们克隆了黄色黏球菌的一个双组分调节系统(phoR2-phoP2),其中phoR2编码组氨酸激酶,phoP2编码应答调节因子。通过使用phoP2作为探针,鉴定并分离出了第二个系统phoR3-phoP3。这两个系统非常相似,组氨酸激酶全长蛋白的同源性为52%,应答调节因子的同源性为64%。两种激酶的预测结构表明它们锚定在膜上,传感结构域位于周质空间,激酶结构域位于细胞质中。应答调节因子(PhoP2和PhoP3)在位于C端区域的效应结构域中表现出典型的DNA结合蛋白的螺旋-环-螺旋基序。对两个单缺失突变体和一个双缺失突变体的研究表明,这些系统参与发育过程。突变体子实体排列不紧密,形成松散扁平的聚集体,其中一些黏孢子不能正常重塑,它们仍保持为伸长的细胞。这些系统还参与了不依赖镁的酸性和中性磷酸酶的表达,这些磷酸酶在发育过程中表达。中性磷酸酶基因尤其依赖PhoP3。PhoP2和PhoP3都不调节碱性磷酸酶和pph1基因的表达。