Muñoz-Dorado J, Inouye S, Inouye M
Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School at Rutgers, Piscataway 08854.
Cell. 1991 Nov 29;67(5):995-1006. doi: 10.1016/0092-8674(91)90372-6.
PCR reactions were carried out on the genomic DNA of M. xanthus, a soil bacterium capable of differentiation to form fruiting bodies, using oligonucleotides representing highly conserved regions of eukaryotic protein serine/threonine kinases. A gene (pkn1) thus cloned contains an ORF of 693 amino acid residues whose amino-terminal domain shows significant sequence similarity with the catalytic domain of eukaryotic protein serine/threonine kinases. The pkn1 gene was overexpressed in E. coli, and the gene product has been found to be autophosphorylated at both serine and threonine residues. The expression of pkn1 is developmentally regulated to start immediately before spore formation. When pkn1 is deleted, differentiation starts prematurely, resulting in poor spore production. These results indicate that the protein serine/threonine kinase plays an important role in the onset of proper differentiation.
利用代表真核生物蛋白丝氨酸/苏氨酸激酶高度保守区域的寡核苷酸,对能分化形成子实体的土壤细菌黄色粘球菌的基因组DNA进行了聚合酶链反应(PCR)。由此克隆出的一个基因(pkn1)包含一个693个氨基酸残基的开放阅读框,其氨基末端结构域与真核生物蛋白丝氨酸/苏氨酸激酶的催化结构域具有显著的序列相似性。pkn1基因在大肠杆菌中过表达,并且发现该基因产物在丝氨酸和苏氨酸残基上都能发生自身磷酸化。pkn1的表达受到发育调控,在孢子形成前立即开始。当pkn1缺失时,分化会过早开始,导致孢子产量低下。这些结果表明,蛋白丝氨酸/苏氨酸激酶在正常分化的起始过程中起重要作用。