US3 protein is one of the most important protein kinases in duck plague virus (DPV), which causes serious economic losses in waterfowl industry due to its high morbidity and mortality. In this article, it illustrates that DPV US3 protein kinase phosphorylated viral protein UL47 and regulated UL47 subcellular localization. US3 protein interacted and co-localized with UL47 in the cytoplasm, while UL47 protein was observed in the nucleus in lack of US3 protein or its kinase activity. The change of UL47 localization caused by US3 protein kinase was independent of their interaction. Finally, the six sites of DPV UL47 at Thr29, Ser30, Ser42, Thr47, Ser161, and Thr775 were identified as the phosphorylation targets of US3 protein, which are different from the related reports of other alphaherpesviruses. This study was published in Frontiers in Microbiology (IF=6.064), and titled as “Duck plague virus US3 protein kinase phosphorylates UL47 and regulates the subcellular localization of UL47”.DOI: 10.3389/fmicb.2022.876820.