{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T06:08:50Z","timestamp":1790834930254,"version":"4.1.0"},"reference-count":63,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,5,22]],"date-time":"2024-05-22T00:00:00Z","timestamp":1716336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100012807","name":"Midwestern University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100012807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Coxiella burnetii<\/jats:italic>\n                      is a gram-negative obligate intracellular bacterium and a zoonotic pathogen that causes human Q fever. The lack of effective antibiotics and a licensed vaccine for\n                      <jats:italic>Coxiella<\/jats:italic>\n                      in the U.S. warrants further research into\n                      <jats:italic>Coxiella<\/jats:italic>\n                      pathogenesis. Within the host cells,\n                      <jats:italic>Coxiella<\/jats:italic>\n                      replicates in an acidic phagolysosome-like vacuole termed\n                      <jats:italic>Coxiella<\/jats:italic>\n                      -containing vacuole (CCV). Previously, we have shown that the CCV pH is critical for\n                      <jats:italic>Coxiella<\/jats:italic>\n                      survival and that the\n                      <jats:italic>Coxiella<\/jats:italic>\n                      Type 4B secretion system regulates CCV pH by inhibiting the host endosomal maturation pathway. However, the trafficking pattern of the \u2018immature\u2019 endosomes in\n                      <jats:italic>Coxiella<\/jats:italic>\n                      - infected cells remained unclear.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We transfected HeLa cells with GFP-tagged Rab proteins and subsequently infected them with mCherry-\n                      <jats:italic>Coxiella<\/jats:italic>\n                      to visualize Rab protein localization. Infected cells were immunostained with anti-Rab antibodies to confirm the Rab localization to the CCV, to quantitate Rab11a and Rab35- positive CCVs, and to quantitate total recycling endosome content of infected cells. A dual-hit siRNA mediated knockdown combined with either immunofluorescent assay or an agarose-based colony-forming unit assay were used to measure the effects of Rab11a and Rab35 knockdown on CCV area and\n                      <jats:italic>Coxiella<\/jats:italic>\n                      intracellular growth.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      The CCV localization screen with host Rab proteins revealed that recycling endosome-associated proteins Rab11a and Rab35 localize to the CCV during infection, suggesting that CCV interacts with host recycling endosomes during maturation. Interestingly, only a subset of CCVs were Rab11a or Rab35-positive at any given time point. Quantitation of Rab11a\/Rab35-positive CCVs revealed that while Rab11a interacts with the CCV more at 3 dpi, Rab35 is significantly more prevalent at CCVs at 6 dpi, suggesting that the CCV preferentially interacts with Rab11a and Rab35 depending on the stage of infection. Furthermore, we observed a significant increase in Rab11a and Rab35 fluorescent intensity in\n                      <jats:italic>Coxiella<\/jats:italic>\n                      -infected cells compared to mock, suggesting that\n                      <jats:italic>Coxiella<\/jats:italic>\n                      increases the recycling endosome content in infected cells. Finally, siRNA-mediated knockdown of Rab11a and Rab35 resulted in significantly smaller CCVs and reduced\n                      <jats:italic>Coxiella<\/jats:italic>\n                      intracellular growth, suggesting that recycling endosomal Rab proteins are essential for CCV expansion and bacterial multiplication.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      Our data, for the first time, show that the CCV dynamically interacts with host recycling endosomes for\n                      <jats:italic>Coxiella<\/jats:italic>\n                      intracellular survival and potentially uncovers novel host cell factors essential for\n                      <jats:italic>Coxiella<\/jats:italic>\n                      pathogenesis.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2024.1394019","type":"journal-article","created":{"date-parts":[[2024,5,22]],"date-time":"2024-05-22T00:38:05Z","timestamp":1716338285000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Coxiella burnetii-containing vacuoles interact with host recycling endosomal proteins Rab11a and Rab35 for vacuolar expansion and bacterial growth"],"prefix":"10.3389","volume":"14","author":[{"given":"Brooke A.","family":"Hall","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristen E.","family":"Senior","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolle T.","family":"Ocampo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dhritiman","family":"Samanta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,5,22]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2017.00397","article-title":"Legionella effector ankX disrupts host cell endocytic recycling in a phosphocholination-dependent manner","volume":"7","author":"Allgood","year":"2017","journal-title":"Front. 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