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Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium

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 wa...

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Autores principales: Figueroa-Cuilan, Wanda M., Irazoki, Oihane, Feeley, Marissa, Smith, Erika, Nguyen, Trung, Cava, Felipe, Goley, Erin D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10295487/
https://www.ncbi.nlm.nih.gov/pubmed/37017481
http://dx.doi.org/10.1091/mbc.E23-01-0023
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author Figueroa-Cuilan, Wanda M.
Irazoki, Oihane
Feeley, Marissa
Smith, Erika
Nguyen, Trung
Cava, Felipe
Goley, Erin D.
author_facet Figueroa-Cuilan, Wanda M.
Irazoki, Oihane
Feeley, Marissa
Smith, Erika
Nguyen, Trung
Cava, Felipe
Goley, Erin D.
author_sort Figueroa-Cuilan, Wanda M.
collection PubMed
description 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.
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spelling pubmed-102954872023-08-16 Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium Figueroa-Cuilan, Wanda M. Irazoki, Oihane Feeley, Marissa Smith, Erika Nguyen, Trung Cava, Felipe Goley, Erin D. Mol Biol Cell Articles 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. The American Society for Cell Biology 2023-06-01 /pmc/articles/PMC10295487/ /pubmed/37017481 http://dx.doi.org/10.1091/mbc.E23-01-0023 Text en © 2023 Figueroa-Cuilan et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial-Share Alike 4.0 International Creative Commons License.
spellingShingle Articles
Figueroa-Cuilan, Wanda M.
Irazoki, Oihane
Feeley, Marissa
Smith, Erika
Nguyen, Trung
Cava, Felipe
Goley, Erin D.
Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title_full Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title_fullStr Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title_full_unstemmed Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title_short Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
title_sort quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10295487/
https://www.ncbi.nlm.nih.gov/pubmed/37017481
http://dx.doi.org/10.1091/mbc.E23-01-0023
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