Marcos A T, Gutiérrez S, Díez B, Fernández F J, Oguiza J A, Martín J F
Department of Ecology, Genetics and Microbiology, Faculty of Biology, University of León, Spain.
Gene. 1995 Feb 3;153(1):41-8. doi: 10.1016/0378-1119(94)00759-l.
Three genes (hrd) homologous to the rpoD gene of Escherichia coli, that encode sigma factor-like proteins, have been cloned from DNA of the candicidin-producing strain Streptomyces griseus IMRU 3570. They are located in different regions of the chromosome. Sequence analysis showed that the first one is analogous to the hrdB gene of S. coelicolor. The second showed high similarity to the hrdD gene of S. coelicolor and S. aureofaciens and is linked, as in S. coelicolor, to a N-acetyltransferase-encoding gene (nat) distantly related to the pat and bar genes that encode resistance to bialafos. The third showed no close homology with other known hrd genes from actinomycetes and has been named hrdT. Functional domains in the three S. griseus Hrd proteins are highly conserved in relation to those of the sigma 70 protein family. Northern analysis showed that hrdB is expressed as a 1.9-kb transcript during active growth in phosphate-rich medium, but it is less efficiently transcribed under sporulation conditions (phosphate-starved) or after a heat-shock treatment. Two other shorter transcripts of 1.2 and 0.7 kb were also detected with the same probe. The hrdD gene is transcribed as a single 1.1-kb transcript under sporulation conditions following nutritional shiftdown and, to a lower extent, during growth conditions in phosphate-rich medium. The hrdT gene is weakly transcribed (1.5-kb RNA) under all conditions tested. The hrd-encoded sigma factors probably recognize actinomycetes promoters (SEP type) with E. coli-like consensus sequences.
已从产杀假丝菌素的灰色链霉菌IMRU 3570的DNA中克隆出三个与大肠杆菌rpoD基因同源的基因(hrd),它们编码类σ因子蛋白。这三个基因位于染色体的不同区域。序列分析表明,第一个基因类似于天蓝色链霉菌的hrdB基因。第二个基因与天蓝色链霉菌和金色链霉菌的hrdD基因高度相似,并且与编码bialafos抗性的pat和bar基因远缘相关的N - 乙酰转移酶编码基因(nat)相连,如同在天蓝色链霉菌中一样。第三个基因与放线菌中其他已知的hrd基因没有密切同源性,已被命名为hrdT。灰色链霉菌的三种Hrd蛋白中的功能域与σ70蛋白家族的功能域高度保守。Northern分析表明,hrdB在富含磷酸盐的培养基中活跃生长期间以1.9 kb的转录本形式表达,但在孢子形成条件下(磷酸盐饥饿)或热休克处理后转录效率较低。用同一探针还检测到另外两个较短的1.2 kb和0.7 kb转录本。hrdD基因在营养转换后的孢子形成条件下转录为单一的1.1 kb转录本,在富含磷酸盐的培养基中的生长条件下转录程度较低。hrdT基因在所有测试条件下转录较弱(1.5 kb RNA)。hrd编码的σ因子可能识别具有类似大肠杆菌共有序列的放线菌启动子(SEP型)。