An SRR1 domain-containing protein is required for efficient Orsay virus replication in Caenorhabditis elegans

Viruses depend on their hosts for completing their life cycle, and a better understanding of virus replication can inform therapeutic strategies. Using the Orsay virus-Caenorhabditis elegans experimental platform, we identified by a forward genetic screen the host gene Y55F3BL.4 (renamed viro-9) as a novel host factor critical for Orsay virus replication. Three distinct mutations of viro-9 each resulted in a >1,000-fold reduction in Orsay viral load, demonstrating a pro-viral function of viro-9. viro-9 had no previously described function in C. elegans, but in the absence of viral infection, deletion of the viro-9 locus by CRISPR/Cas9 led to a reduction in brood size and a shortened lifespan. VIRO-9 contains a sensitivity to red light reduced (SRR1) protein domain. While SRR1 domains are present in diverse organisms, including plants, yeast, and mammals, little is known about their function. The Caenorhabditis briggsae ortholog of viro-9, CBG23913, can functionally complement the C. elegans viro-9 defect, demonstrating that the pro-viral function of the SRR1 domain is conserved over at least 80 million years of evolution. Furthermore, we identified three conserved amino acid residues within the SRR1 domain that are required for Orsay virus infection. This study provides the first insights into amino acids necessary for functionality of the SRR1 domain and demonstrates the essential role of viro-9 in virus infection.IMPORTANCEHost factors required for viral replication could serve as therapeutic targets for various viral species. The Caenorhabditis elegans-Orsay virus experimental system offers a platform for identifying genes important for virus infection in nematodes that may also be important for human-infecting viruses. We determined that viro-9, a previously uncharacterized gene in C. elegans containing the SRR1 domain, is required for Orsay virus replication. The related gene in Caenorhabditis briggsae, a relative of C. elegans that diverged about 80 million years ago, can substitute for viro-9, demonstrating that this protein's ability to promote virus replication is functionally conserved. Because SRR1 domain-containing proteins exist in nematodes, fungi, Drosophila, plants, and mammals, including humans, these proteins could be important for facilitating virus infection in other organisms as well.