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In this study, we performed microarray analysis to detect genes with differential expression between the \u0394<jats:italic>rfbE<\/jats:italic>mutant and wild-type strain S2308. Interestingly, acid shock protein 24 gene (<jats:italic>asp24<\/jats:italic>) expression was significantly upregulated in the \u0394<jats:italic>rfbE<\/jats:italic>mutant compared to S2308, as confirmed by quantitative reverse transcription-PCR (qRT-PCR) and Western blotting. Further studies using additional strains indicated that the upregulation of<jats:italic>asp24<\/jats:italic>occurred only in rough mutants with disrupted O-antigen export system components, including the ATP-binding protein gene<jats:italic>rfbE<\/jats:italic>(<jats:italic>bab1_0542<\/jats:italic>) and the permease gene<jats:italic>rfbD<\/jats:italic>(<jats:italic>bab1_0543<\/jats:italic>), while the \u0394<jats:italic>wboA<\/jats:italic>rough mutant (which lacks an O-antigen synthesis-related glycosyltransferase) and the RB51 strain (a vaccine strain with the rough phenotype) showed no significant changes in<jats:italic>asp24<\/jats:italic>expression compared to S2308. In addition, abolishing the intracellular O-antigen synthesis of the \u0394<jats:italic>rfbE<\/jats:italic>mutant by deleting the<jats:italic>wboA<\/jats:italic>gene (thereby creating the \u0394<jats:italic>rfbE<\/jats:italic>\u0394<jats:italic>wboA<\/jats:italic>double-knockout strain) recovered<jats:italic>asp24<\/jats:italic>expression. These results indicated that<jats:italic>asp24<\/jats:italic>upregulation is associated with intracellular O-antigen synthesis and accumulation but not with the bacterial rough phenotype. Further studies indicated that<jats:italic>asp24<\/jats:italic>upregulation in the \u0394<jats:italic>rfbE<\/jats:italic>mutant was associated neither with bacterial adherence and invasion nor with cellular necrosis on RAW264.7 macrophages. However, proper expression of the<jats:italic>asp24<\/jats:italic>gene favors intracellular survival of<jats:named-content content-type=\"genus-species\">Brucella<\/jats:named-content>in RAW264.7 cells and HeLa cells during an infection. This study reveals a novel mechanism for<jats:italic>asp24<\/jats:italic>upregulation in<jats:named-content content-type=\"genus-species\">B. abortus<\/jats:named-content>mutants.<\/jats:p>","DOI":"10.1128\/iai.01765-14","type":"journal-article","created":{"date-parts":[[2014,4,22]],"date-time":"2014-04-22T02:51:20Z","timestamp":1398135080000},"page":"2840-2850","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":15,"title":["Mechanism of Asp24 Upregulation in Brucella abortus Rough Mutant with a Disrupted O-Antigen Export System and Effect of Asp24 in Bacterial Intracellular Survival"],"prefix":"10.1128","volume":"82","author":[{"given":"Mingxing","family":"Tian","sequence":"first","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Qu","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"},{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiangan","family":"Han","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chan","family":"Ding","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaohui","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daxin","family":"Peng","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Yangzhou University, Yangzhou, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shengqing","family":"Yu","sequence":"additional","affiliation":[{"name":"Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(00)00165-X"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-090110-102905"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11515-012-1196-0"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1612.091081"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2011.00054"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2012.00002"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.022622699"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1017\/S1462399407000543"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1286-4579(02)01604-0"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-5822.2005.00609.x"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2008.01.005"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-5-13"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetmic.2007.11.023"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2004.12.003"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.46.2.384-388.1984"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(87)80702-0"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.8.3830-3835.1999"},{"key":"e_1_3_3_19_2","doi-asserted-by":"crossref","first-page":"677","DOI":"10.2460\/ajvr.1996.57.05.677","article-title":"Protection of BALB\/c mice against homologous and heterologous species of Brucella by rough strain vaccines derived from Brucella melitensis and Brucella suis biovar 4","volume":"57","author":"Winter AJ","year":"1996","unstructured":"WinterAJSchurigGGBoyleSMSriranganathanNBevinsJSEnrightFMElzerPHKopecJD. 1996. 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