Luong Thanh T, Ouyang Shu, Bush Kelly, Lee Chia Y
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
J Bacteriol. 2002 Jul;184(13):3623-9. doi: 10.1128/JB.184.13.3623-3629.2002.
The cap1 genes are required for the synthesis of type 1 capsular polysaccharide (CP1) in Staphylococcus aureus. We previously showed that the cap1 locus was associated with a discrete genetic element in S. aureus M. In this report, we defined the boundaries of the cap1 element by comparing its restriction pattern to that of a corresponding region from the CP1-negative strain Becker. The element was located in the SmaI-G chromosomal fragment of the standard mapping strain NCTC8325. The sequences of the entire cap1 element and the flanking regions were determined. We found that there were two additional cap1 genes not previously identified. The cap1 operon was located in a staphylococcal cassette chromosome (SCC) element similar to the resistance island SCCmec recently described for methicillin resistance in S. aureus. Notably, the SCCcap1 element was located at the same insertion site as all the SCCmec elements in the staphylococcal chromosome. The excision of SCCcap1 could be demonstrated only in the presence of the recombinase genes from an SCCmec element, verifying that SCCcap1 is a genuine SCC element but defective in mobilization. A novel enterotoxin gene, whose transcript was detected by Northern blotting, was found next to the SCCcap1 locus. We propose that the enterotoxin gene and SCCcap1 were inserted into this locus at the juxtaposition by independent events. Sequence comparison revealed numerous DNA rearrangements and mutations in SCCcap1 and the left flanking region, suggesting that the SCCcap1 had been inserted at the SCC attC site a long time ago. In addition, most genes in this region were incomplete, with the exception of the 15 cap1 genes, implying that the cap1 genes confer a survival advantage on strain M.
cap1基因是金黄色葡萄球菌合成1型荚膜多糖(CP1)所必需的。我们之前表明,cap1基因座与金黄色葡萄球菌M中的一个离散遗传元件相关。在本报告中,我们通过将其限制性图谱与CP1阴性菌株贝克尔相应区域的图谱进行比较,确定了cap1元件的边界。该元件位于标准图谱菌株NCTC8325的SmaI - G染色体片段中。测定了整个cap1元件及其侧翼区域的序列。我们发现有两个之前未鉴定出的额外cap1基因。cap1操纵子位于一个葡萄球菌盒式染色体(SCC)元件中,类似于最近描述的与金黄色葡萄球菌耐甲氧西林相关的耐药岛SCCmec。值得注意的是,SCCcap1元件位于葡萄球菌染色体中与所有SCCmec元件相同的插入位点。只有在存在来自SCCmec元件的重组酶基因时,才能证明SCCcap1的切除,这证实SCCcap1是一个真正的SCC元件,但在移动性方面存在缺陷。在SCCcap1基因座旁边发现了一个新的肠毒素基因,其转录本通过Northern印迹法检测到。我们提出肠毒素基因和SCCcap1是通过独立事件并列插入到这个基因座中的。序列比较揭示了SCCcap1及其左侧侧翼区域存在大量DNA重排和突变,表明SCCcap1很久以前就已插入到SCC attC位点。此外,该区域的大多数基因是不完整的,除了15个cap1基因,这意味着cap1基因赋予了菌株M生存优势。