Damjanovic Marlen, Kharat Arun S, Eberhardt Alice, Tomasz Alexander, Vollmer Waldemar
Microbial Genetics, University of Tübingen, Tübingen, Germany.
J Bacteriol. 2007 Oct;189(19):7105-11. doi: 10.1128/JB.00681-07. Epub 2007 Jul 27.
Streptococcus pneumoniae has an absolute nutritional requirement for choline, and the choline molecules are known to incorporate exclusively into the cell wall and membrane teichoic acids of the bacterium. We describe here the isolation of a mutant of strain R6 in which a single G-->T point mutation in the gene tacF (formerly designated spr1150) is responsible for generating a choline-independent phenotype. The choline-independent phenotype could be transferred to the laboratory strain R6 and to the encapsulated strain D39 by genetic transformation with a PCR product or with a plasmid carrying the mutated tacF gene. The tacF gene product belongs to the protein family of polysaccharide transmembrane transporters (flippases). A model is presented in which TacF is required for the transport of the teichoic acid subunits across the cytoplasmic membrane. According to this model, wild-type TacF has a strict specificity for choline-containing subunits, whereas the TacF present in the choline-independent mutant strain is able to transport both choline-containing and choline-free teichoic acid chains. The proposed transport specificity of parental-type TacF for choline-containing subunits would ensure the loading of the cell wall with teichoic acid chains decorated with choline residues, which appear to be essential for the virulence of this pathogen.
肺炎链球菌对胆碱有绝对的营养需求,已知胆碱分子仅掺入该细菌的细胞壁和膜磷壁酸中。我们在此描述了R6菌株突变体的分离,其中基因tacF(以前称为spr1150)中的单个G→T点突变导致产生胆碱非依赖型表型。通过用PCR产物或携带突变tacF基因的质粒进行遗传转化,胆碱非依赖型表型可以转移到实验室菌株R6和包膜菌株D39中。tacF基因产物属于多糖跨膜转运蛋白(翻转酶)家族。本文提出了一个模型,其中TacF是磷壁酸亚基跨细胞质膜运输所必需的。根据该模型,野生型TacF对含胆碱的亚基具有严格的特异性,而胆碱非依赖型突变菌株中的TacF能够运输含胆碱和不含胆碱的磷壁酸链。亲本型TacF对含胆碱亚基的拟议运输特异性将确保细胞壁加载有装饰有胆碱残基的磷壁酸链,这似乎对该病原体的毒力至关重要。