Functional analysis of the interaction between viral protein m and tropomyosin 3

• Grant number: 19/11310-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2019
• End date: July 31, 2020
• Field of knowledge: Biological Sciences - Microbiology
• Principal Investigator: Armando Morais Ventura
• Grantee: Beatriz Domenici de Oliveira
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Human Respiratory Syncytial Virus (HRSV) is the most important pathological agent of the respiratory tract, affecting mainly newborns and infants. It is estimated that there are 34 million new cases per year, resulting in about 200,000 deaths. Because of the large number of hospitalizations due to HRSV infection, its socioeconomic impact is considered to be a major public health burden. There is no approved vaccine and the treatments available in the market are inefficient, demonstrating the importance of the characterization of new therapeutic targets. It was verified in the laboratory previously that the viral proteins phosphoprotein (P) and matrix protein (M) interact with tropomyosin isoform 3 (TPM3) in the infected cell. Data from the literature show that the HRSV infection exerts a strong interference on the intracellular actin location, a phenomenon that we propose to be related to the interaction of TPM3 with M and P, leading to the rearrangement of microfilaments. To test this hypothesis, we aim to functionally analyze the interaction of viral protein M with TPM3. To this end, we will analyze the replication of the virus in cells treated with a drug that promotes destabilization of the microfilaments or in silenced cells for TPM3 with shRNA. In addition, we aim to construct and analyze the functionality of FLAG-TPM3 to confirm interactions with viral proteins using methodologies such as immunoprecipitation and immunofluorescence.