Gliding motility and cytadherence in Mycoplasma penetrans

性状（由申请方提供）：大多数具有极化细胞形态的支原体种属表现出滑动运动性，这是一个与宿主细胞粘附（细胞粘附）密不可分的过程，但分子机制不确定。此外，不同的系统发育组的支原体似乎粘附和滑行的不同机制。反式支原体是一种主要在HIV阳性患者中发现的生物体，它似乎通过促进HIV感染的细胞类型的增殖来加强AIDS的进展，它也表现出极性形态，并且已知会钻入宿主上皮细胞。然而，其与宿主细胞相互作用的分子基础是完全未知的，其基因组序列表明没有其他生物的细胞粘附蛋白的同源物。初步数据表明，M.与其他极化支原体一样，反式支原体在体外表现出滑行运动性，其速度与其菌落吸附红细胞（红细胞吸附）的程度有关，这表明在该种属中，细胞粘附和滑行也相关。我们将通过延时显微电影摄影术描述快速滑动菌株和慢速滑动菌株的滑动运动性，包括旨在确定M的能量来源的实验。横向滑翔。我们将表征已经存在的血细胞吸附（HA）突变体以及我们产生和选择的新HA突变体的滑行能力，以进一步研究M.是的。我们将使用二维聚丙烯酰胺凝胶电泳和质谱鉴定蛋白质是不存在的或改变这些突变体作为候选人参与cytadherence和/或滑行运动。我们将提高抗这些蛋白质的抗血清，开始研究他们如何在这些过程中发挥作用，使用的抗体作为功能的抑制剂，并作为工具来分析亚细胞定位和稳定性的亲本菌株和突变株。这些实验的结果将揭示一种新的细胞推进分子机制，最终可能导致合理的治疗设计，也可能有助于纳米技术机器的发展。

DESCRIPTION (provided by applicant): Most mycoplasma species with polarized cell morphologies exhibit gliding motility, a process inextricably intertwined with adherence to host cells (cytadherence) but of uncertain molecular mechanism. Moreover, different phylogenetic groups of mycoplasmas appear to adhere and glide by different mechanisms. Mycoplasma penetrans, an organism found principally in HIV-positive patients in whom it appears to potentiate the progress of AIDS by promoting proliferation of the type of cell that HIV infects, also exhibits polar morphology and is known to burrow into host epithelial cells. However, the molecular basis for its interaction with host cells is entirely unknown, its genome sequence indicating an absence of homologs of cytadherence proteins of other organisms. Preliminary data indicate that M. penetrans, like other polarized mycoplasmas, exhibits gliding motility in vitro, its speed relating to the degree to which its colonies adsorb erythrocytes (hemadsorb), suggesting that in this species, cytadherence and gliding are also related. We will characterize the gliding motility of a fast-gliding strain and a slow-gliding strain by time-lapse microcinematography with respect to speed and optimal gliding conditions, including experiments aimed at identifying the energy source for M. penetrans gliding. We will characterize the gliding ability of both already-existing hemadsorption (HA) mutants as well as new HA mutants that we generate and select in order to further investigate the relationship between cytadherence and gliding motility in M. penetrans. We will use two-dimensional polyacrylamide gel electrophoresis and mass spectrometry to identify proteins that are absent or altered in these mutants as candidates for involvement in cytadherence and/or gliding motility. We will raise antisera against these proteins to begin studies of how they function in these processes, using the antibodies both as inhibitors of function and as tools to analyze subcellular localization and stability in both the parent strain and the mutant strains. The results of these experiments will reveal a novel molecular mechanism for cellular propulsion that might ultimately lead to rational design of therapeutics and might also be useful in the development of nanotechnological machines.