000075520 001__ 75520
000075520 005__ 20200117221655.0
000075520 0247_ $$2doi$$a10.1186/s13567-018-0583-1
000075520 0248_ $$2sideral$$a107954
000075520 037__ $$aART-2018-107954
000075520 041__ $$aeng
000075520 100__ $$aPerez-Etayo, L.
000075520 245__ $$aThe CO2-dependence of Brucella ovis and Brucella abortus biovars is caused by defective carbonic anhydrases
000075520 260__ $$c2018
000075520 5060_ $$aAccess copy available to the general public$$fUnrestricted
000075520 5203_ $$aBrucella bacteria cause brucellosis, a major zoonosis whose control requires efficient diagnosis and vaccines. Identification of classical Brucella spp. has traditionally relied on phenotypic characterization, including surface antigens and 5-10% CO2 necessity for growth (CO2-dependence), a trait of Brucella ovis and most Brucella abortus biovars 1-4 strains. Although molecular tests are replacing phenotypic methods, CO2-dependence remains of interest as it conditions isolation and propagation and reflects Brucella metabolism, an area of active research. Here, we investigated the connection of CO2-dependence and carbonic anhydrases (CA), the enzymes catalyzing the hydration of CO2 to the bicarbonate used by anaplerotic and biosynthetic carboxylases. Based on the previous demonstration that B. suis carries two functional CAs (CAI and CAII), we analyzed the CA sequences of CO2-dependent and -independent brucellae and spontaneous mutants. The comparisons strongly suggested that CAII is not functional in CO2-dependent B. abortus and B. ovis, and that a modified CAII sequence explains the CO2-independent phenotype of spontaneous mutants. Then, by mutagenesis and heterologous plasmid complementation and chromosomal insertion we proved that CAI alone is enough to support CO2-independent growth of B. suis in rich media but not of B. abortus in rich media or B. suis in minimal media. Finally, we also found that insertion of a heterologous active CAII into B. ovis reverted the CO2-dependence but did not alter its virulence in the mouse model. These results allow a better understanding of central aspects of Brucella metabolism and, in the case of B. ovis, provide tools for large-scale production of diagnostic antigens and vaccines.
000075520 536__ $$9info:eu-repo/grantAgreement/ES/DGA/A13-17D$$9info:eu-repo/grantAgreement/ES/MINECO/AGL2014-58795-C4-1-R$$9info:eu-repo/grantAgreement/ES/MINECO/AGL2014-58795-C4-3-R
000075520 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000075520 590__ $$a3.117$$b2018
000075520 591__ $$aVETERINARY SCIENCES$$b4 / 141 = 0.028$$c2018$$dQ1$$eT1
000075520 592__ $$a0.124$$b2018
000075520 593__ $$aVeterinary (miscellaneous)$$c2018$$dQ4
000075520 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000075520 700__ $$ade Miguel, M.J.
000075520 700__ $$aConde-Alvarez, R.
000075520 700__ $$0(orcid)0000-0002-7970-4929$$aMuñoz, P.M.
000075520 700__ $$aKhames, M.
000075520 700__ $$aIriarte, M.
000075520 700__ $$aMoriyon, I.
000075520 700__ $$aZuniga-Ripa, A.
000075520 773__ $$g49 (2018), 85 [12 pp]$$pVet. res.$$tVeterinary Research$$x0928-4249
000075520 8564_ $$s1322263$$uhttps://zaguan.unizar.es/record/75520/files/texto_completo.pdf$$yVersión publicada
000075520 8564_ $$s101905$$uhttps://zaguan.unizar.es/record/75520/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000075520 909CO $$ooai:zaguan.unizar.es:75520$$particulos$$pdriver
000075520 951__ $$a2020-01-17-22:10:40
000075520 980__ $$aARTICLE