Identification and characterization of putative Aeromonas spp. T3SS effectors

Autor(es)

Rangel, Luiz Thiberio ^{[1, 2, 3, 4]} ; Marden, Jeremiah ^[3] ; Colston, Sophie ^{[3, 5]} ; Setubal, Joao Carlos ^{[1, 4]} ; Graf, Joerg ^{[3, 6]} ; Gogarten, Johann Peter ^{[3, 6]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo - Brazil

^[2] MIT, Dept Earth Atmospher & Planetary Sci, Cambridge, MA - USA

^[3] Univ Connecticut, Dept Mol & Cell Biol, Storrs, CT 06269 - USA

^[4] Univ Sao Paulo, Interunidades Bioinformat, Sao Paulo - Brazil

^[5] US Naval Res Lab, Ctr Bio Mol Sci & Engn, Washington, DC - USA

^[6] Univ Connecticut, Inst Syst Genom, Storrs, CT 06269 - USA

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 14, n. 6 JUN 4 2019.
• Citações Web of Science: 0

Resumo

The genetic determinants of bacterial pathogenicity are highly variable between species and strains. However, a factor that is commonly associated with virulent Gram-negative bacteria, including many Aeromonas spp., is the type 3 secretion system (T3SS), which is used to inject effector proteins into target eukaryotic cells. In this study, we developed a bioinformatics pipeline to identify T3SS effector proteins, applied this approach to the genomes of 105 Aeromonas strains isolated from environmental, mutualistic, or pathogenic contexts and evaluated the cytotoxicity of the identified effectors through their heterologous expression in yeast. The developed pipeline uses a two-step approach, where candidate Aeromonas gene families are initially selected using Hidden Markov Model (HMM) profile searches against the Virulence Factors DataBase (VFDB), followed by strict comparisons against positive and negative control datasets, greatly reducing the number of false positives. This approach identified 21 Aeromonas T3SS likely effector families, of which 8 represent known or characterized effectors, while the remaining 13 have not previously been described in Aeromonas. We experimentally validated our in silico findings by assessing the cytotoxicity of representative effectors in Saccharomyces cerevisiae BY4741, with 15 out of 21 assayed proteins eliciting a cytotoxic effect in yeast. The results of this study demonstrate the utility of our approach, combining a novel in silico search method with in vivo experimental validation, and will be useful in future research aimed at identifying and authenticating bacterial effector proteins from other genera. (AU)

• Processo FAPESP: 14/23975-0 - Transferência horizontal de genes ligados a adaptação a novos nichos em procariotos
• Beneficiário: Luiz Thibério Lira Diniz Rangel
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 12/17196-2 - Redes Filogenômicas e Transferencia Lateral de Genes
• Beneficiário: Luiz Thibério Lira Diniz Rangel
• Modalidade de apoio: Bolsas no Brasil - Doutorado