Kim H J, Calcutt M J, Schmidt F J, Chater K F
John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom.
J Bacteriol. 2000 Mar;182(5):1313-20. doi: 10.1128/JB.182.5.1313-1320.2000.
Candidate partitioning genes (parA and parB) for the linear chromosome of Streptomyces coelicolor were identified by DNA sequencing in a series of seven genes located between rnpA and trxA near the chromosomal replication origin. The most likely translation start point of parB overlapped the parA stop codon, suggestive of coregulation, and transcription analysis suggested that the two genes formed an operon. Deletion of part of parB had no effect on the growth or appearance of colonies but caused a deficiency in DNA partitioning during the multiple septation events involved in converting aerial hyphae into long chains of spores. At least 13% of spore compartments failed to inherit the normal DNA allocation. The same phenotype was obtained with a deletion removing a segment of DNA from both parA and parB. Reinforcing the idea of a special role for the par locus during sporulation, the stronger of two parAB promoters was greatly upregulated at about the time when sporulation septation was maximal in colonies. Three copies of a 14-bp inverted repeat (GTTTCACGTGAAAC) were found in or near the parAB genes, and at least 12 more identical copies were identified within 100 kb of oriC from the growing genome sequence database. Only one perfect copy of the 14-bp sequence was present in approximately 5 Mb of sequence available from the rest of the genome. The 14-bp sequence was similar to sequences identified as binding sites for Spo0J, a ParB homologue from Bacillus subtilis believed to be important for DNA partitioning (D. C.-H. Lin and A. D. Grossman, Cell 92:675-685, 1998). One of these sites encompassed the transcription start point of the stronger parA promoter.
通过对天蓝色链霉菌线性染色体的候选分区基因(parA和parB)进行DNA测序,在位于染色体复制起点附近的rnpA和trxA之间的一系列七个基因中鉴定出了它们。parB最可能的翻译起始点与parA终止密码子重叠,提示存在共调控,转录分析表明这两个基因形成一个操纵子。删除parB的部分区域对菌落的生长或外观没有影响,但在气生菌丝转化为长链孢子所涉及的多次隔膜形成过程中,导致DNA分区出现缺陷。至少13%的孢子间隔未能继承正常的DNA分配。删除parA和parB的一段DNA也获得了相同的表型。在菌落中孢子形成隔膜最多的时候,parAB两个启动子中较强的一个在此时大幅上调,这进一步强化了par基因座在孢子形成过程中具有特殊作用的观点。在parAB基因内或附近发现了三个14碱基对的反向重复序列(GTTTCACGTGAAAC),并且从不断增长的基因组序列数据库中发现在oriC的100 kb范围内至少还有12个相同的拷贝。在基因组其余部分约5 Mb的序列中仅存在一个14碱基对序列的完美拷贝。该14碱基对序列与被鉴定为枯草芽孢杆菌的ParB同源物Spo0J的结合位点的序列相似,据信Spo0J对DNA分区很重要(D. C.-H. Lin和A. D. Grossman,《细胞》92:675 - 685,1998)。其中一个位点包含较强的parA启动子的转录起始点。