A Zinc-dependent metalloproteinase in the intracellular adaptation of Brucella abortus in macrophages

BioRxiv : the Preprint Server for Biology
L. A. GomezA. Onate


Brucella abortus is a pathogen that survives in macrophages. Several virulence factors participate in this process, including the open reading frame (ORF) BAB1_0270 codifying of a Zinc-dependent metalloproteinase. Here, its contribution in the process of intracellular adaptation was analyzed by infecting RAW264.7 macrophages with the mutant B. abortus {Delta}270 strain. Results showed that this Zinc-dependent metalloproteinase is a cytoplasmic protein that conforms an operon with a transcriptional regulator, which may constitute a type II toxin-antitoxin system. Functionally, this Zinc-dependent metalloproteinase participated neither in the adherence nor the initial intracellular traffic of B. abortus in macrophages. Nevertheless, its deletion significantly increased the co-localization of B. abortus {Delta}270 with phagolysosomal cathepsin D, reducing both its co-localization with calnexin, present in endoplasmic reticulum derived vesicles, and its intracellular replication within macrophages. Besides, B. abortus {Delta}270-infected macrophages produced significantly higher levels of TNF-, IL-6, CD80 and CD86 than B. abortus 2308, even when several genes involved in virulence ( vjbR , hutC , bvrR , virB1 ) were up-regulated in...Continue Reading

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