biostudies-literatureKang BAgricultural Science and Technology Innovation ProgramChina Agriculture Research System of MOF and MARANational Natural Science Foundation of China3431https://www.ebi.ac.uk/biostudies/studies/S-EPMC11644367biostudies-literatureUnknown13(23)Potyviral P3 is involved in viral replication, movement, and pathogenicity; however, its biochemical function is unknown. In this study, the P3 of the zucchini yellow mosaic virus (ZYMV) interacted with ClBBD, a protein with high ortholog bifunctional nuclease activity, in watermelon. The binding site was shown via yeast two-hybrid screening and BiFC assay to be located at the N-terminus of P3 rather than P3N-PIPO. ClBBD localized predominantly to the chloroplast and plasma membrane. ZYMV P3 was also present in the nucleus and cytoplasm as aggregates. When co-expressed with P3 in tobacco, ClBBD formed aggregates with P3 in the cytoplasm. The knockdown of <i>ClBBD</i> using the VIGS vector pV190 and challenge with ZYMV revealed a positive correlation between viral accumulation and ClBBD expPlants (Basel, Switzerland)A Bifunctional Nuclease Promotes the Infection of Zucchini Yellow Mosaic Virus in Watermelon by Targeting P3.PMC11644367CARS-25CAAS-ASTIP-2022-ZFRI-09U21A2022931701942Kang BLiu LLiu MPeng BLiang ZZang YGu QLiu FWu HfalseA Bifunctional Nuclease Promotes the Infection of Zucchini Yellow Mosaic Virus in Watermelon by Targeting P3.Potyviral P3 is involved in viral replication, movement, and pathogenicity; however, its biochemical function is unknown. In this study, the P3 of the zucchini yellow mosaic virus (ZYMV) interacted with ClBBD, a protein with high ortholog bifunctional nuclease activity, in watermelon. The binding site was shown via yeast two-hybrid screening and BiFC assay to be located at the N-terminus of P3 rather than P3N-PIPO. ClBBD localized predominantly to the chloroplast and plasma membrane. ZYMV P3 was also present in the nucleus and cytoplasm as aggregates. When co-expressed with P3 in tobacco, ClBBD formed aggregates with P3 in the cytoplasm. The knockdown of <i>ClBBD</i> using the VIGS vector pV190 and challenge with ZYMV revealed a positive correlation between viral accumulation and ClBBD exp2024-01-01T00:00:00Z2024 Dec2026-04-23T03:24:52.413Z2025-04-04T01:54:38.134ZS-EPMC116443673968322410.3390/plants13233431