Characterization of the cell interactome of proteins E, M and N of the new Coronavirus of 2019 (SARS-CoV-2)

• Grant number: 20/09310-6
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): September 01, 2020
• Effective date (End): June 30, 2022
• Field of knowledge: Health Sciences - Nutrition - Nutrition Biochemistry
• Principal researcher: Fernando Moreira Simabuco
• Grantee: Érika Pereira Zambalde
• Home Institution: Faculdade de Ciências Aplicadas (FCA). Universidade Estadual de Campinas (UNICAMP). Limeira , SP, Brazil
• Associated research grant: 20/05346-6 - Interaction studies between cellular proteins and viral proteins of the new coronavirus 2019 (SARS-CoV-2), AP.R

Abstract

SARS-CoV-2 causes COVID19, characterized by fever, cough, fatigue, and severe cases of pneumonia, having affected more than 10 million people worldwide so far. Three of the virus' structural proteins, proteins E, M, and N, are responsible for the viral particle assembly, among other processes. Protein N has a high affinity for viral RNA, forming the viral nucleocapsid. The membrane proteins E and M are responsible for the morphology of the virion and for interactions with the S protein, which recognizes the cellular receptor, and with the viral nucleocapsid. The replication process of coronaviruses is highly regulated, involving different actions of the non-structural and structural proteins of the virus. However, few is known about the cellular proteins involved in the assembly of the viral particles of the Coronaviruses. This project aims to identify the interactome of the E, M, and N proteins of SARS-CoV-2 in human cells, understanding their molecular interactions with cellular proteins and the cellular pathways that benefit this virus to complete its replication cycle. For that, we will perform the optimization of the genes that encode E, M, and N for expression in human cells, followed by immunoprecipitation and characterization by mass spectrometry of interacting cellular proteins. Once validated, such interactions will be challenged with pharmacological interventions in vitro, to verify their potential to block or interfere with viral replication. These compounds will finally be tested in an in vitro culture model of human cells infected with SARS-CoV-2, analyzing viral load, cytopathic effect, and molecular changes in the cells. The present research project aims, therefore, to contribute to a better understanding of the molecular and cellular mechanisms associated with the replication of SARS-CoV-2 in human cells and to rationalize potential future therapies against COVID-19. (AU)