Study of the involvement of ESCRT machinery in the assembly and budding of zika virus in human glioblastoma cells

• Grant number: 19/20193-4
• Support type: Scholarships in Brazil - Master
• Effective date (Start): March 01, 2020
• Effective date (End): November 30, 2021
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal researcher: Luis Lamberti Pinto da Silva
• Grantee: Milena Barrocali de Araújo Melo
• Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 14/02438-6 - Studies with Bunyaviridae that produce human disease, AP.TEM

Abstract

Zika virus (ZIKV) is a flavivirus whose main transmitting agent are Aedes family mosquitoes. Despite its relevance in public health and reports in the literature correlating this arbovirus with severe clinical manifestations leading to central nervous system (CNS) impairment, the molecular mechanisms involved in ZIKV assembly and externalization are poorly elucidated. In this context, studies on the intracellular dynamics of ZIKV, particularly regarding the participation of host cell factors in viral propagation, are scarce. The present work aims to investigate and characterize the participation of ESCRT (Endosomal Sorting Complex Required for Transport) machinery in ZIKV replication. Therefore, it will be investigated whether the expression of HRS (ESCRT-0), TSG101 (ESCRT-I), CHMP4 (ESCRT III) subunits and the accessory protein Alix are necessary for ZIKV replication. Preliminary data from our group suggest that TSG101 protein plays an important role in ZIKV replication in U-251 cells, however further experiments will be conducted to confirm these findings and understand at what stage of the viral replicative cycle, TSG101 activity is important. Another objective is to determine whether ZIKV proteins physically interact with ESCRTs. We will also investigate the TSG101 domain involved in the interaction, and will use the GFP-trap technique to explore other candidate proteins that may interact with viral factors. Studies such as this, which aim to elucidate how ZIKV uses the host cellular machinery, are essential for understanding the viral replicative cycle at the molecular level, contributing to the elaboration of strategies directed to antiviral therapies. (AU)