IBBM   21076
INSTITUTO DE BIOTECNOLOGIA Y BIOLOGIA MOLECULAR
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Proteomic analysis of Bordetella bronchiseptica with high intracellular c-di-GMP concentration
Autor/es:
SISTI, FEDERICO; FERNANDEZ, JULIETA
Lugar:
Buenos Aires
Reunión:
Simposio; 11th International Bordetella Symposium; 2016
Resumen:
Objective: We have already described second messenger c-di-GMP (cdG) role in motility and biofilm formation through overexpression of the diguanilate cyclase BdcA (BB_RS17940). Over expressing BdcA in B. bronchiseptica triggers high c-di-GMP levels. The objective of this work was to identify proteins positively regulated by cdG in BdcA over expressing strain.Material and methods: We performed quantitative comparative proteomic to find proteins involved in cdG control of known and possibly new phenotypes. Cytosolic fractions of Bb-BdcA and wild type (WT) were compared.Results: More than 850 proteins were identified by this method. Proteins detected range in different categories; 35,35% are metabolism related, 15% related to translation process, 6,3% are involved in protein folding or other postranscription process. Interestingly, we found 79 (11%) proteins predicted to be located at the periplasm and 8 proteins involved in cell wall biosynthesis. Some of the known virulence factors were present in both strains; fimbrae, FHA, cyA, PRN and four proteins of the type three secretio system, bopN, bcsQ, bopD and bscW. We detected 24 proteins significantly different expressed. A group of seven proteins were only present in Bb-BdcA in all biological replica performed belonging to metabolism, secA independent secretion systems, and unknown categories. Seventeen proteins were significantly repressed by BdcA. Among them a putative hidrolase was absent in Bb-BdcA.Conclusion: This work presented the quadrupole Orbitrap mass spectrometer technique to analyze bacterial proteomes.Many proteins showed significantly differences expression profiles, encouraging us to elucidate their role in cdG regulon in Bordetellla