{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,17]],"date-time":"2025-11-17T17:23:20Z","timestamp":1763400200245,"version":"3.41.2"},"reference-count":45,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2024,6,20]],"date-time":"2024-06-20T00:00:00Z","timestamp":1718841600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/doi.org\/10.1128\/ASMCopyrightv2"},{"start":{"date-parts":[[2024,6,20]],"date-time":"2024-06-20T00:00:00Z","timestamp":1718841600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["R35GM130320"],"award-info":[{"award-number":["R35GM130320"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000199","name":"U.S. Department of Agriculture","doi-asserted-by":"crossref","award":["USDA-NIFA 7002581"],"award-info":[{"award-number":["USDA-NIFA 7002581"]}],"id":[{"id":"10.13039\/100000199","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["R21AI148132, R01AI172822"],"award-info":[{"award-number":["R21AI148132, R01AI172822"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2024,6,20]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Bacteria rely on DNA methylation for restriction-modification systems and epigenetic control of gene expression. Here, we use direct detection of methylated bases by nanopore sequencing to monitor global DNA methylation in Alphaproteobacteria, where use of this technique has not yet been reported. One representative of this order,\n              <jats:italic>Caulobacter crescentus<\/jats:italic>\n              , relies on DNA methylation to control cell cycle progression, but it is unclear whether other members of this order, such as\n              <jats:italic>Brucella abortus<\/jats:italic>\n              , depend on the same systems. We addressed these questions by first measuring CcrM-dependent DNA methylation in\n              <jats:italic>Caulobacter<\/jats:italic>\n              and showing excellent correlation between nanopore-based detection and previously published results. We then directly measure the impact of Lon-mediated CcrM degradation on the epigenome, verifying that loss of Lon results in pervasive methylation. We also show that the AlkB demethylase has no global impact on DNA methylation during normal growth. Next, we report on the global DNA methylation in\n              <jats:italic>B. abortus<\/jats:italic>\n              for the first time and find that CcrM-dependent methylation is reliant on Lon but impacts the two chromosomes differently. Finally, we explore the impact of the MucR transcription factor, known to compete with CcrM methylation, on the\n              <jats:italic>Brucella<\/jats:italic>\n              methylome and share the results with a publicly available visualization package. Our work demonstrates the utility of nanopore-based sequencing for epigenome measurements in Alphaproteobacteria and reveals new features of CcrM-dependent methylation in a zoonotic pathogen.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                DNA methylation plays an important role in bacteria, maintaining genome integrity and regulating gene expression. We used nanopore sequencing to directly measure methylated bases in\n                <jats:italic>Caulobacter crescentus<\/jats:italic>\n                and\n                <jats:italic>Brucella abortus<\/jats:italic>\n                . In\n                <jats:italic>Caulobacter<\/jats:italic>\n                , we showed that stabilization of the CcrM methyltransferase upon loss of the Lon protease results in prolific methylation and discovered that the putative methylase AlkB is unlikely to have a global physiological effect. We measured genome-wide methylation in\n                <jats:italic>Brucella<\/jats:italic>\n                for the first time, revealing a similar role for CcrM in cell-cycle methylation but a more complex regulation by the Lon protease than in Caulobacter. Finally, we show how the virulence factor MucR impacts DNA methylation patterns in\n                <jats:italic>Brucella<\/jats:italic>\n                .\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/jb.00083-24","type":"journal-article","created":{"date-parts":[[2024,5,9]],"date-time":"2024-05-09T13:00:22Z","timestamp":1715259622000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Comparison of CcrM-dependent methylation in\n            <i>Caulobacter crescentus<\/i>\n            and\n            <i>Brucella abortus<\/i>\n            by nanopore sequencing"],"prefix":"10.1128","volume":"206","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-6352-0463","authenticated-orcid":false,"given":"Maxwell","family":"Campbell","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ian Scott","family":"Barton","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, South Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2101-6928","authenticated-orcid":true,"suffix":"II","given":"R. Martin","family":"Roop","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Brody School of Medicine, East Carolina University, Greenville, South Carolina, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6733-1478","authenticated-orcid":true,"given":"Peter","family":"Chien","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, Massachusetts, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2008.02.012"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.57.2.434-450.1993"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3435"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1994.tb01313.x"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2008.00159.x"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0708112104"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/cdf490"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.10.12.1532"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1319315110"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkt1352"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1006499"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0092-8674(02)01133-9"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrestox.5b00522"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.10.3139-3145.1997"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja3010094"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2000.01726.x"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.182.12.3482-3489.2000"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02201-23"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600927"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.6.2874"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-08289-9"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1906119116"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14500"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2020.07.011"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.177.7.1662-1669.1995"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms5366"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms5081"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01097-12"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0014198"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.3844805"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2019.01.010"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens9110942"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/0882-4010(92)90065-v"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00255-10"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bty191"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btab705"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-021-01147-4"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-023-02910-3"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4184"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btq033"},{"key":"e_1_3_4_43_2","unstructured":"R Core Team. 2021. 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