Role of RNA modifications in coronaviruses replication

National Institute Of Environmental Health Sciences

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Abstract

SARS-CoV-2, the virus responsible for the current pandemic, belongs to the coronavirus family. These viruses have a polyadenylated RNA genome that replicates within the infected cells. The poly(A) tail of the virus is critical for viral RNA integrity and changes in length during the course of the infection. However, the molecular pathways controlling viral RNA synthesis and processing are poorly understood. Coronaviruses encode for several RNA processing enzymes, but they also depend on host proteins for their replication. We hypothesize that some host RNA binding proteins previously implicated in poly(A) processing of other viruses might contribute to the metabolism of coronaviruses' RNA. During this year, we designed and validated strategies to deplete these proteins from mammalian cells. We will later infect cells depleted of the RNA processing proteins to understand their impact on viral replication. To define the molecular function of the host proteins during infection, we established a sequencing protocol and a computational pipeline to measure the virus' poly(A) tail length. The method also allows us to detect internal RNA modifications, which are believed to be necessary for RNA processing too. By measuring the virus poly(A) tail length and RNA modifications in infected cells depleted of key RNA regulatory proteins, we will be able to dissect the molecular pathways controlling the virus replication.