Mutant analysis reveals a specific requirement for protein P30 in Mycoplasma pnenmoniae gliding motility
Mutant analysis reveals a specific requirement for protein P30 in Mycoplasma pnenmoniae gliding motility
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DOI：
10.1128/jb.187.18.6281-6289.2005
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发表时间：
2005-09-01
影响因子：
3.2
通讯作者：
Krause, DC
中科院分区：
文献类型：
--
作者：
Hasselbring, BM;Jordan, JL;Krause, DC
无细胞壁的原核生物肺炎支原体，长期以来被认为是能够自我复制的最小和最简单的细胞之一，具有明显的细胞极性，其特征在于存在分化的末端细胞器，其功能是粘附于人呼吸道上皮、滑动运动和细胞分裂。红细胞吸附（HA）阴性突变体的表征导致了几种末端细胞器蛋白的鉴定，包括P30，其损失导致发育缺陷和对宿主细胞的粘附性降低，但它们对M.尚未研究肺炎滑动。在这里，我们研究的基础上卫星生长和细胞滑行速度和频率的P30滑行运动的贡献。M. pneumoniae HA突变体II-3缺乏P30是不运动的，但产生截短的P30的HA突变体II-7是运动的，尽管速度比野生型低50倍。产生改变的P30的HA阳性回复突变体II-3R出乎意料地在滑动方面不是完全野生型。突变体II-3与重组野生型和突变体等位基因的互补证实了滑动缺陷和P30丢失或改变之间的相关性。令人惊讶的是，黄色荧光蛋白与P30的C末端融合对细胞滑动速度几乎没有影响，并显著增强HA。最后，虽然HA的定量检查显示这些突变株之间有明确的区别，滑动缺陷并不严格相关的HA表型，所有菌株附着在玻璃在野生型水平。两者合计，这些研究结果表明，P30在滑行运动中的作用，这是不同的，从它的要求在坚持。
The cell-wall-less prokaryote Mycoplasma pneumoniae, long considered among the smallest and simplest cells capable of self-replication, has a distinct cellular polarity characterized by the presence of a differentiated terminal organelle which functions in adherence to human respiratory epithelium, gliding motility, and cell division. Characterization of hemadsorption (HA)-negative mutants has resulted in identification of several terminal organelle proteins, including P30, the loss of which results in developmental defects and decreased adherence to host cells, but their impact on M. pneumoniae gliding has not been investigated. Here we examined the contribution of P30 to gliding motility on the basis of satellite growth and cell gliding velocity and frequency. M. pneumoniae HA mutant II-3 lacking P30 was nonmotile, but HA mutant II-7 producing a truncated P30 was motile, albeit at a velocity 50-fold less than that of the wild type. HA-positive revertant II-3R producing an altered P30 was unexpectedly not fully wild type with respect to gliding. Complementation of mutant II-3 with recombinant wild-type and mutant alleles confirmed the correlation between gliding defect and loss or alteration in P30. Surprisingly, fusion of yellow fluorescent protein to the C terminus of P30 had little impact on cell gliding velocity and significantly enhanced HA. Finally, while quantitative examination of HA revealed clear distinctions among these mutant strains, gliding defects did not correlate strictly with the HA phenotype, and all strains attached to glass at wild-type levels. Taken together, these findings suggest a role for P30 in gliding motility that is distinct from its requirement in adherence.