AUTHOR=de la Garza-García Jorge A. , Ouahrani-Bettache Safia , Lyonnais Sébastien , Ornelas-Eusebio Erika , Freddi Luca , Al Dahouk Sascha , Occhialini Alessandra , Köhler Stephan 

TITLE=Comparative Genome-Wide Transcriptome Analysis of Brucella suis and Brucella microti Under Acid Stress at pH 4.5: Cold Shock Protein CspA and Dps Are Associated With Acid Resistance of B. microti

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.794535

DOI=10.3389/fmicb.2021.794535

ISSN=1664-302X

ABSTRACT=<p>Brucellae are facultative intracellular coccobacilli causing brucellosis, one of the most widespread bacterial zoonosis affecting wildlife animals, livestock and humans. The genus <italic>Brucella</italic> comprises classical and atypical species, such as <italic>Brucella suis</italic> and <italic>Brucella microti</italic>, respectively. The latter is characterized by increased metabolic activity, fast growth rates, and extreme acid resistance at pH 2.5, suggesting an advantage for environmental survival. In addition, <italic>B. microti</italic> is more acid-tolerant than <italic>B. suis</italic> at the intermediate pH of 4.5. This acid-resistant phenotype of <italic>B. microti</italic> may have major implications for fitness in soil, food products and macrophages. Our study focused on the identification and characterization of acid resistance determinants of <italic>B. suis</italic> and <italic>B. microti</italic> in Gerhardt’s minimal medium at pH 4.5 and 7.0 for 20 min and 2 h by comparative RNA-Seq-based transcriptome analysis, validated by RT-qPCR. Results yielded a common core response in both species with a total of 150 differentially expressed genes, and acidic pH-dependent genes regulated specifically in each species. The identified core response mechanisms comprise proton neutralization or extrusion from the cytosol, participating in maintaining physiological intracellular pH values. Differential expression of 441 genes revealed species-specific mechanisms in <italic>B. microti</italic> with rapid physiological adaptation to acid stress, anticipating potential damage to cellular components and critical energy conditions. Acid stress-induced genes encoding cold shock protein CspA, pseudogene in <italic>B. suis</italic>, and stress protein Dps were associated with survival of <italic>B. microti</italic> at pH 4.5. <italic>B. suis</italic> response with 284 specifically regulated genes suggested increased acid stress-mediated protein misfolding or damaging, triggering the set-up of repair strategies countering the consequences rather than the origin of acid stress and leading to subsequent loss of viability. In conclusion, our work supports the hypothesis that increased acid stress resistance of <italic>B. microti</italic> is based on selective pressure for the maintenance of functionality of critical genes, and on specific differential gene expression, resulting in rapid adaptation.</p>