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Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, United States.
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, United States.
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, United States.
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2023 (Engelska)Ingår i: Molecular Biology of the Cell, ISSN 1059-1524, E-ISSN 1939-4586, Vol. 34, nr 7, artikel-id ar69Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

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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American Society for Cell Biology (ASCB) , 2023. Vol. 34, nr 7, artikel-id ar69
Nationell ämneskategori
Cell- och molekylärbiologi
Identifikatorer
URN: urn:nbn:se:umu:diva-209113DOI: 10.1091/mbc.E23-01-0023PubMedID: 37017481Scopus ID: 2-s2.0-85159238537OAI: oai:DiVA.org:umu-209113DiVA, id: diva2:1763937
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This article is distributed by The American Societyfor Cell Biology under license from the author(s). Two months after publication itis available to the public under an Attribution–Noncommercial-Share Alike 4.0 International Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/4.0)

Tillgänglig från: 2023-06-08 Skapad: 2023-06-08 Senast uppdaterad: 2023-07-19Bibliografiskt granskad

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Irazoki, OihaneCava, Felipe

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Irazoki, OihaneCava, Felipe
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Molekylär Infektionsmedicin, Sverige (MIMS)Umeå Centre for Microbial Research (UCMR)Institutionen för molekylärbiologi (Medicinska fakulteten)
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Molecular Biology of the Cell
Cell- och molekylärbiologi

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