Screening bacterial effectors and human virus proteins in yeast to identify host factors driving tombusvirus RNA recombination: a role for autophagy and membrane phospholipid content

Positive-strand (+)RNA viruses replicate in the cytosol of infected cells by exploiting cellular proteins and resources that frequently lead to diseases. Virus replication results in the generation of viral RNA recombinants that contribute to the emergence of new viral variants and adaptation to new hosts. The authors expressed <i>Legionella</i> bacterium effector proteins, SARS-CoV-2 and human metapneumovirus proteins in yeast to test their effects on tomato bushy stunt virus (TBSV) RNA recombination. This novel approach revealed that <i>Legionella</i> effectors and heterologous viral proteins target shared host factors with TBSV, including the autophagy pathway. <i>In vitro</i> approach revealed that the pro-recombination role of co-opted autophagy is to provide abundant phospholipids for viral replication. SARS-CoV-2 nucleocapsid protein and human metapneumovirus M2-1 protein are shown to enhance TBSV RNA replication and recombination by protecting the viral RNAs from host Xrn1 5´-3´ exoribonuclease in yeast. Thus, the TBSV/yeast system can be used as a cellular system sensor to find new functions of heterologous viral proteins.