McCallum K L, Whitfield C
Department of Microbiology, University of Guelph, Ontario, Canada.
Infect Immun. 1991 Feb;59(2):494-502. doi: 10.1128/iai.59.2.494-502.1991.
In Klebsiella pneumoniae, the ability to synthesize large amounts of capsular polysaccharide is an important correlate of virulence. We report the cloning of rcsA from K. pneumoniae serotype O1:K20 and demonstrate that rcsA is involved in the expression of the K antigen capsule. We have determined the nucleotide sequence for the rcsA gene from K. pneumoniae K20 and shown it to be identical to the sequence reported previously for rcsA from strain K21 (Allen et al., J. Gen. Microbiol. 133:331-340, 1987). Southern hybridization results indicate that this gene is widely distributed among different Klebsiella K serotypes. When cloned into Escherichia coli K-12, the K. pneumoniae rcsA gene caused a mucoid phenotype, resulting from the activation of colanic acid synthesis. Activation of colanic acid synthesis was not dependent on growth at low temperatures (less than or equal to 30 degrees C). The K. pneumoniae rcsA gene complemented E. coli K-12 rcsA mutations but could not complement defects in rcsB, suggesting that RcsA may be functionally homologous in these bacteria. The cloned rcsA gene also complemented a defect in nonmucoid strain K20 derivatives that normally produced only trace amounts of K20 antigen and were unable to assemble a wild-type capsular structure. Mutants that were K20-deficient were not complemented. The K antigen capsule of K. pneumoniae therefore joins a growing list of polysaccharide-synthetic systems in which "RcsA-like" proteins are involved.
在肺炎克雷伯菌中,合成大量荚膜多糖的能力是毒力的一个重要相关因素。我们报道了从肺炎克雷伯菌O1:K20血清型中克隆rcsA,并证明rcsA参与K抗原荚膜的表达。我们已经确定了肺炎克雷伯菌K20的rcsA基因的核苷酸序列,并表明它与先前报道的K21菌株的rcsA序列相同(Allen等人,《普通微生物学杂志》133:331 - 340,1987)。Southern杂交结果表明该基因在不同的克雷伯菌K血清型中广泛分布。当克隆到大肠杆菌K - 12中时,肺炎克雷伯菌的rcsA基因导致了黏液样表型,这是由结肠酸合成的激活引起的。结肠酸合成的激活不依赖于低温(小于或等于30℃)下的生长。肺炎克雷伯菌的rcsA基因弥补了大肠杆菌K - 12的rcsA突变,但不能弥补rcsB的缺陷,这表明RcsA在这些细菌中可能在功能上是同源的。克隆的rcsA基因也弥补了非黏液样菌株K20衍生物中的缺陷,这些衍生物通常只产生微量的K20抗原,并且无法组装野生型荚膜结构。缺乏K20的突变体没有得到弥补。因此,肺炎克雷伯菌的K抗原荚膜加入了越来越多涉及“类RcsA”蛋白的多糖合成系统的列表中。