Orthoreovirus outer-fiber proteins are substrates for SUMO-conjugating enzyme Ubc9.
Reoviruses are potential anticancer agents due to their ability to induce cell death in tumor cells. Grass carp reovirus (GCRV) is one of the best characterized models on reovirus pathogenesis in vitro. However, there is little known about how SUMOylation affects reovirus pathogenesis. The SUMO conjugating enzyme 9 (Ubc9) determines the targets of SUMOylation. Here, the protein interactions between reovirus outer fiber proteins, specifically GCRV-104 VP55, and Ubc9 were probed using a yeast two-hybrid system. The N-terminal coiled-coil domain of VP55, containing a single lysine residue, was responsible for the interaction between VP55 and Ubc9 in yeast. In solid phase binding assays, a single amino acid mutation (K87R) prevented Ubc9 from binding to VP55. Overexpression of Ubc9 enhanced GCRV-104 infection efficiency, and knockdown of Ubc9 in CIK cells inhibited viral replication, which suggested that Ubc9 was a proviral factor. Furthermore, Ubc9 was shown to bind outer fiber proteins from type II GCRV, avian reovirus and mammalian reovirus in yeast. To our knowledge, this is the first study to show that Ubc9 binds to reovirus outer-fiber proteins and likely contributes to efficient orthoreovirus replication. These results suggest that SUMOylation modifications could be targeted to improve the therapeutic efficacy of oncolytic reovirus.
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Aquatic Genetic Resources of Ministry of Aquaculture, Shanghai Ocean University, Shanghai, PR China.
Fei
F
Yu
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Aquatic Genetic Resources of Ministry of Aquaculture, Shanghai Ocean University, Shanghai, PR China.
Hao
H
Wang
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Aquatic Genetic Resources of Ministry of Aquaculture, Shanghai Ocean University, Shanghai, PR China.
Longlong
L
Wang
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Aquatic Genetic Resources of Ministry of Aquaculture, Shanghai Ocean University, Shanghai, PR China.
Liqun
L
Lu
D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps / Q000821:virology; D005393:Fish Diseases / Q000378:metabolism / Q000821:virology; D006470:Hemorrhage / Q000378:metabolism / Q000821:virology; D018165:Orthoreovirus / Q000737:chemistry / Q000378:metabolism; D010802:Phylogeny; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D012088:Reoviridae Infections / Q000378:metabolism / Q000473:pathology / Q000662:veterinary; D016415:Sequence Alignment; D058207:Sumoylation; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*
D014764:Viral Proteins; D044763:Ubiquitin-Conjugating Enzymes
SUMOylation; Ubc9; outer-fiber protein; reovirus