Kormanec J, Farkasovský M
Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava.
Nucleic Acids Res. 1993 Aug 11;21(16):3647-52. doi: 10.1093/nar/21.16.3647.
In previous experiments, Streptomyces aureofaciens has been shown to contain four genes hrdA, hrdB, hrdD, and hrdE, encoding polypeptides very similar to principal sigma factors of RNA polymerase. Two apparent tandem promoters were identified for each of the hrdA, hrdB and hrdD genes by S1 nuclease mapping using RNA prepared of S. aureofaciens in various developmental stages. Under all the conditions studied, tandem promoters of each gene differed significantly in their respective strengths. Transcription from the hrd promoters depended on developmental stage. While hrdB is transcribed from both promoters in all developmental stages, both tandem promoters of the hrdD gene are active only in vegetative stage and transcription of the hrdA tandem promoters temporally correlates with the aerial mycelium formation. In addition to a promoter, hrdB-P2, which lies upstream of the open reading frame, the hrdB gene, proposed to encode functional principal factor, appeared to contain at least one internal promoter, hrdB-P1. Activity of all promoters was consistent with S1 mapping experiments after insertion of promoter-bearing DNA fragments to promoter-probe vectors pIJ486 and pARC1. The results implicate temporally different expression of the hrd genes during the differentiation of S. aureofaciens.
在先前的实验中,已表明金色链霉菌含有四个基因hrdA、hrdB、hrdD和hrdE,它们编码的多肽与RNA聚合酶的主要σ因子非常相似。通过使用在不同发育阶段制备的金色链霉菌RNA进行S1核酸酶图谱分析,为hrdA、hrdB和hrdD基因分别鉴定出两个明显的串联启动子。在所有研究条件下,每个基因的串联启动子在各自的强度上有显著差异。hrd启动子的转录取决于发育阶段。虽然hrdB在所有发育阶段都从两个启动子转录,但hrdD基因的两个串联启动子仅在营养阶段活跃,hrdA串联启动子的转录与气生菌丝体的形成在时间上相关。除了位于开放阅读框上游的启动子hrdB - P2外,推测编码功能性主要因子的hrdB基因似乎还包含至少一个内部启动子hrdB - P1。将携带启动子的DNA片段插入启动子探针载体pIJ486和pARC1后,所有启动子的活性与S1图谱分析实验结果一致。这些结果表明在金色链霉菌分化过程中hrd基因存在时间上不同的表达。