{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T05:56:46Z","timestamp":1787032606384,"version":"build-2736575974"},"posted":{"date-parts":[[2026,7,13]]},"group-title":"Microbiology","reference-count":70,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T00:00:00Z","timestamp":1783900800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["R35GM124733"],"award-info":[{"award-number":["R35GM124733"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R35GM131762"],"award-info":[{"award-number":["R35GM131762"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["F32AI174818"],"award-info":[{"award-number":["F32AI174818"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01AI177619"],"award-info":[{"award-number":["R01AI177619"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006710","name":"Wayne State University","doi-asserted-by":"crossref","award":["Career Chair to JMS"],"award-info":[{"award-number":["Career Chair to JMS"]}],"id":[{"id":"10.13039\/100006710","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,13]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Bacterial gene expression depends on the coordinated regulation of RNA synthesis, processing, and decay. The conserved RNA degradosome scaffold, Ribonuclease E (RNase E), assembles into phase-separated bacterial ribonucleoprotein bodies (BR-bodies) through its C-terminal intrinsically disordered region (IDR). This IDR also scaffolds recruitment of degradosome client proteins that carry out post-transcriptional gene regulation. Whether BR-bodies regulate virulence programs to support infection, however, is largely undefined. We show that RNase E of the intracellular pathogen\n                  <jats:italic>Brucella ovis<\/jats:italic>\n                  forms RNA-dependent condensates in vivo and phase-separates with RNA in vitro, with the IDR necessary and sufficient for BR-body assembly. Deleting the IDR (\n                  <jats:italic>rne<\/jats:italic>\n                  (\u0394IDR)) did not impair growth but sensitized\n                  <jats:italic>Brucella<\/jats:italic>\n                  to host-relevant oxidative and cell-envelope stressors. Transcriptome-wide profiling that simultaneously resolved mRNA decay and processing revealed that BR-bodies primarily accelerate mRNA turnover while stabilizing a distinct subset of transcripts. Notably, BR-bodies control the processing and levels of\n                  <jats:italic>virB<\/jats:italic>\n                  type IV secretion system (T4SS) mRNA and the turnover of its key activators, linking condensate-based RNA regulation to a core virulence pathway. Consistent with\n                  <jats:italic>virB<\/jats:italic>\n                  dysregulation, the\n                  <jats:italic>rne<\/jats:italic>\n                  (\u0394IDR) mutant was severely attenuated in mammalian macrophages. To test whether phase separation is sufficient for BR-body function, we replaced the\n                  <jats:italic>B. ovis<\/jats:italic>\n                  IDR with the highly divergent\n                  <jats:italic>Caulobacter crescentus<\/jats:italic>\n                  IDR. This chimera assembled BR-bodies but rescued fitness incompletely, fully restoring oxidative-stress resistance but not cell-envelope stress resistance or intracellular fitness. BR-bodies therefore link RNA metabolism to\n                  <jats:italic>Brucella<\/jats:italic>\n                  stress resistance and infection, and their full function requires both phase separation and additional native IDR-specific activities such as degradosome interactions.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    Biomolecular condensates are non-membrane bound organelles that organize biological processes across all domains of life, but their role in bacterial infection is not well characterized. We show that\n                    <jats:italic>Brucella<\/jats:italic>\n                    , an intracellular bacterial pathogen and causative agent of the disease brucellosis, relies on biomolecular condensates called BR-bodies to control RNA stability and regulate gene expression. Without BR-bodies,\n                    <jats:italic>Brucella<\/jats:italic>\n                    is sensitized to host-relevant chemical stresses, cannot properly regulate essential infection machinery, and has severely diminished fitness within mammalian host cells. These results indicate that control of RNA levels by biomolecular condensates is required for\n                    <jats:italic>Brucella<\/jats:italic>\n                    survival within animal hosts, providing evidence that phase separation is a fundamental mechanism underlying\n                    <jats:italic>Brucella<\/jats:italic>\n                    adaptation to the hostile environment encountered during infection.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.11.737834","type":"posted-content","created":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T05:55:18Z","timestamp":1784008518000},"source":"Crossref","is-referenced-by-count":1,"title":["BR-bodies link RNA homeostasis to stress tolerance and intracellular fitness in\n                  <i>Brucella<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5201-2279","authenticated-orcid":false,"given":"Kaveendya S.","family":"Mallikaarachchi","sequence":"first","affiliation":[{"name":"Department of Biology, Indiana University, Bloomington, IN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7780-6355","authenticated-orcid":false,"given":"Rosemary","family":"Northcote","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4930-5401","authenticated-orcid":false,"given":"Thomas","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4067-6331","authenticated-orcid":false,"given":"Melene A.","family":"Alakavuklar","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9609-2701","authenticated-orcid":false,"given":"Vincent","family":"Lawal","sequence":"additional","affiliation":[{"name":"Department of Biology, Indiana University, Bloomington, IN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vidhyadhar","family":"Nandana","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Wayne State University, Detroit, MI, USA"},{"name":"Nuclear Dynamics and Cancer Program, Cancer Epigenetics Institute, Fox Chase Cancer Center, Philadelphia, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0612-5029","authenticated-orcid":false,"given":"Aretha","family":"Fiebig","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5728-5882","authenticated-orcid":false,"given":"Jared M.","family":"Schrader","sequence":"additional","affiliation":[{"name":"Department of Biology, Indiana University, Bloomington, IN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1727-322X","authenticated-orcid":false,"given":"Sean","family":"Crosson","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, 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