Gaurivaud Patrice, Persson Anja, Grand Dominique Le, Westberg Joakim, Solsona Michel, Johansson Karl-Erik, Poumarat François
UMR Mycoplasmoses des Ruminants, AFSSA-Site de Lyon, 31 av Tony Garnier, FR-69364 Lyon Cedex 07, France.
Microbiology (Reading). 2004 Dec;150(Pt 12):4009-22. doi: 10.1099/mic.0.27247-0.
Intraclonal antigenic variation in pathogenic mycoplasma species is considered an important feature of host-pathogen interaction. Such intraclonal protein variation was observed for the interaction of Mycoplasma mycoides subsp. mycoides Small Colony, the agent of contagious bovine pleuropneumonia, with mAb 3F3. Colony immunostaining allows the definition of 3F3 ON- and 3F3 OFF-type variants, which revert at low frequency. Targets of mAb 3F3 were shown to be surface located, and resided on multiple polypeptides in the 58-68 kDa size range. Phage display and a genomic database were combined to determine the gene encoding the proteins recognized by mAb 3F3. A gene encoding the putative permease of the glucose phosphotransferase system was identified. Genome sequence analysis of strain PG1 revealed two highly similar copies of this gene, resulting from duplication of the chromosomal region carrying the gene. Southern blot analysis demonstrated the presence of this duplication in almost every African strain tested, but not in European strains. DNA analysis revealed that ON/OFF switching is governed by a base substitution occurring upstream of the coding region for the 3F3 epitope. This event generates a stop codon that results in the premature termination of the PtsG protein.
致病性支原体物种的克隆内抗原变异被认为是宿主-病原体相互作用的一个重要特征。在丝状支原体丝状亚种小菌落型(即传染性牛胸膜肺炎病原体)与单克隆抗体3F3的相互作用中观察到了这种克隆内蛋白质变异。菌落免疫染色可定义3F3 ON型和3F3 OFF型变体,它们会以低频率回复突变。单克隆抗体3F3的靶标显示位于表面,存在于大小在58-68 kDa范围内的多种多肽上。将噬菌体展示和基因组数据库相结合,以确定编码被单克隆抗体3F3识别的蛋白质的基因。鉴定出一个编码葡萄糖磷酸转移酶系统推定通透酶的基因。PG1菌株的基因组序列分析揭示了该基因的两个高度相似的拷贝,这是由携带该基因的染色体区域重复所致。Southern印迹分析表明,几乎每个测试的非洲菌株中都存在这种重复,但欧洲菌株中没有。DNA分析显示,ON/OFF转换受3F3表位编码区上游发生的碱基替换控制。这一事件产生一个终止密码子,导致PtsG蛋白过早终止。