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The role of D2 subunit of vacuolar ATPase protein (ATP6V0d2) in the biogenesis of vacuoles formed by Coxiella burnetii infection in raw 264.7 macrophages

Grant number: 17/14070-1
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): October 01, 2017
Effective date (End): August 31, 2019
Field of knowledge:Biological Sciences - Parasitology
Principal researcher:Renato Arruda Mortara
Grantee:Ana Carolina Benites Badaró
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Coxiella burnetii is an obligate intracellular pathogen and etiological agent of the zoonosisknown as Q fever. It is capable of infects several cell types in vitro and harboring in acidic pH (Coxiella-containing vacuoles, CCV) intracellular vacuoles that provide favorable conditions for its multiplication and metabolism. Phagocytosis is a defense mechanism that involves the formation of phagolysosomes for the degradation of pathogens. However, some microorganisms such as C. burnetii are able to form vacuoles and survive within these acidic structures. An important characteristic of mature vacuoles formed by pathogen infections is the intense activity of proteins responsible for vacuolar acidification, vacuolar ATPases (V-ATPases). The V-ATPases are organized in V1 domain composed of eight subunits and V0 composed of five subunits, are responsible for the acidification of the vacuoles and are present in the CCVs. Studies with RAW 264.7 macrophages silenced for d2 subunit of the V-ATPases(ATP6V0d2) infected with Leishmania amazonensis demonstrated that these parasitophorous vacuoles, morphologically similar to the CCVs, presented a decrease in its volume. Considering that the studies of C. burnetii infection involving macrophages are extremely important for the understanding of the pathogenicity and spreading of the bacteria, since they mimic the natural infection in vivo, in this project we will evaluate the formation of C. burnetii vacuoles during infection in macrophages RAW 264.7 for d2 subunit of the V-ATPases. We will also investigate whether the bacterial vacuolar growth is dependent on ATP6V0d2, as observed in L. amazonensis infection. (AU)

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