Homerová D, Sevcíková J, Kormanec J
Institute of Molecular Biology, Slovak Academy of Sciences, 845 51 Bratislava, Slovakia.
Folia Microbiol (Praha). 2003;48(4):489-95. doi: 10.1007/BF02931330.
The gene corresponding to the recently identified whiB-paralogous gene wblE in S. coelicolor was found after sequencing the downstream region of the stress-response sporulation-specific sigma-factor gene, sigH, in S. coelicolor A3(2). Sequence analysis has revealed an ORF exhibiting high similarity to sporulation transcription factors WhiB and WhiD. A stable null mutant of the wblE gene was obtained by integrative transformation, via double cross-over. Disruption of the S. coelicolor wblE gene appeared to have no obvious effect on growth, morphology, differentiation, and production of the pigmented antibiotics actinorhodin and undecylprodigiosin. Expression of the wblE gene was investigated during differentiation by S1 nuclease mapping, using RNA prepared from S. coelicolor A3(2) and its isogenic sigF and sigH mutants grown to various developmental stages. A single promoter was identified upstream of the wblE coding region. The wblEp promoter was induced at the beginning of aerial mycelium formation and its activity decreased later in differentiation. No differences in expression of the wblEp promoter were detected in S. coelicolor A3(2) mutants in sigF and sigH genes for sporulation-specific sigma factors. Sequence of the wblEp promoter showed partial similarity to the consensus sequence of the extracytoplasmic sigma factors.
在对天蓝色链霉菌A3(2)中应激反应孢子形成特异性σ因子基因sigH的下游区域进行测序后,发现了与最近鉴定出的天蓝色链霉菌中whiB旁系同源基因wblE相对应的基因。序列分析揭示了一个与孢子形成转录因子WhiB和WhiD具有高度相似性的开放阅读框。通过双交换整合转化获得了wblE基因的稳定缺失突变体。天蓝色链霉菌wblE基因的破坏似乎对生长、形态、分化以及色素抗生素放线紫红素和十一烷基灵菌红素的产生没有明显影响。利用从天蓝色链霉菌A3(2)及其同基因sigF和sigH突变体在不同发育阶段生长所制备的RNA,通过S1核酸酶图谱分析研究了wblE基因在分化过程中的表达。在wblE编码区上游鉴定出一个单一启动子。wblEp启动子在气生菌丝体形成开始时被诱导,其活性在分化后期降低。在孢子形成特异性σ因子的sigF和sigH基因的天蓝色链霉菌A3(2)突变体中未检测到wblEp启动子表达的差异。wblEp启动子的序列与胞外σ因子的共有序列显示出部分相似性。