Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium.
Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium.
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DOI：
10.1091/mbc.e23-01-0023
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发表时间：
2023-06-01
影响因子：
3.3
通讯作者：
Goley, Erin D.
中科院分区：
文献类型：
--
作者：
Figueroa-Cuilan, Wanda M.;Irazoki, Oihane;Feeley, Marissa;Smith, Erika;Nguyen, Trung;Cava, Felipe;Goley, Erin D.
立克次体目的专性细胞内细菌包括重要的人类病原体。然而，我们对立克次体物种生物学的理解受到其专性细胞内生活方式所带来的挑战的限制。为了克服这一障碍，我们开发了方法来评估细胞壁组成，生长和形态的立克次体parkeri，人类病原体斑点热组的立克次体属。细胞壁的分析。parkeri揭示了区别于自由生活的α-变形菌的独特特征。使用一种新的荧光显微镜方法，我们定量R。在活宿主细胞中观察到Parkeri形态，并发现经历细胞分裂的群体比例在感染过程中降低。我们进一步证明了定位荧光融合的可行性，例如，细胞分裂蛋白ZapA，在活R。帕克第一次。为了评估群体生长动力学，我们开发了一种基于成像的测定方法，该方法提高了其他方法的通量和分辨率。最后，我们应用这些工具来定量地证明肌动蛋白同源物MreB是R.帕克里生长和杆形。总体而言，开发了一套高通量定量工具来了解R.帕克里（parkeri）可翻译为其他专性细胞内细菌。
Obligate intracellular bacteria of the order Rickettsiales include important human pathogens. However, our understanding of the biology of Rickettsia species is limited by challenges imposed by their obligate intracellular lifestyle. To overcome this roadblock, we developed methods to assess cell wall composition, growth, and morphology of Rickettsia parkeri, a human pathogen in the spotted fever group of the Rickettsia genus. Analysis of the cell wall of R. parkeri revealed unique features that distinguish it from free-living alphaproteobacteria. Using a novel fluorescence microscopy approach, we quantified R. parkeri morphology in live host cells and found that the fraction of the population undergoing cell division decreased over the course of infection. We further demonstrated the feasibility of localizing fluorescence fusions, for example, to the cell division protein ZapA, in live R. parkeri for the first time. To evaluate population growth kinetics, we developed an imaging-based assay that improves on the throughput and resolution of other methods. Finally, we applied these tools to quantitatively demonstrate that the actin homologue MreB is required for R. parkeri growth and rod shape. Collectively, a toolkit was developed of high-throughput, quantitative tools to understand growth and morphogenesis of R. parkeri that is translatable to other obligate intracellular bacteria.
影响因子：
15.9
作者：
Wyckoff, Timna J.;Taylor, Jennifer A.;Salama, Nina R.
通讯作者：
Salama, Nina R.
影响因子：
5.4
作者：
Lee TK;Huang KC
通讯作者：
Huang KC