title: Phosphorylation of Ser-80 of VP1 and Ser-254 of VP2 is essential for human BK virus propagation in tissue culture.
abstract: BK virus (BKV) infection may cause polyomavirus-associated nephropathy in patients with renal transplantation. Recently, the phosphorylated amino acids on the structural proteins VP1, VP2 and VP3 of BKV have been identified by liquid chromatography-tandem mass spectrometry in our laboratory. In this study, we further analysed the biological effects of these phosphorylation events. Phosphorylation of the BKV structural proteins was demonstrated by [(32)P]orthophosphate labelling in vivo. Site-directed mutagenesis was performed to replace all of the phosphorylated amino acids. The mutated BKV genomes were transfected into Vero cells for propagation analysis. The results showed that expression of the early protein LT and of the late protein VP1 by the mutants VP1-S80A, VP1-S80-133A, VP1-S80-327A, VP1-S80-133-327A and VP2-S254A was abolished. However, propagation of other mutants was similar to that of wild-type BKV. The results suggest that phosphorylation of Ser-80 of VP1 and Ser-254 of VP2 is crucial for BKV propagation.
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan, ROC.|Department of Chemistry and Biochemistry, National Chung Cheng University, Chia-Yi, Taiwan, ROC.|Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan, ROC.
authors.forename: Pei-Lain
authors.initials: PL
authors.lastname: Chen
authors.affiliation: Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan, ROC.|Department of Life Science, National Taiwan Ocean University, Keelung, Taiwan, ROC.
authors.forename: Pang-Hung
authors.initials: PH
authors.lastname: Hsu
authors.affiliation: Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan, ROC.
authors.forename: Chiung-Yao
authors.initials: CY
authors.lastname: Fang
authors.affiliation: Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan, ROC.
authors.forename: Chi-Fang
authors.initials: CF
authors.lastname: Chang
authors.affiliation: Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan, ROC.
authors.forename: Wei-Chih
authors.initials: WC
authors.lastname: Ou
authors.affiliation: Department of Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan, ROC.
authors.forename: Meilin
authors.initials: M
authors.lastname: Wang
authors.affiliation: Institute of Molecular Biology, National Chung Cheng University, Chia-Yi, Taiwan, ROC.
authors.forename: Deching
authors.initials: D
authors.lastname: Chang
mesh_terms: D000818:Animals; D001739:BK Virus / Q000235:genetics / Q000502:physiology*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D007553:Isotope Labeling; D016297:Mutagenesis, Site-Directed; D010761:Phosphorus Radioisotopes / Q000378:metabolism; D010766:Phosphorylation; D012694:Serine / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D010761:Phosphorus Radioisotopes; C062481:VP1 protein, polyomavirus; C442543:VP2 protein, Polyomavirus; D012694:Serine