RT Journal Article SR Electronic(1) A1 Huang, Mingshu A1 Sato, Hiroki A1 Hagiwara, Kyoji A1 Watanabe, Akira A1 Sugai, Akihiro A1 Ikeda, Fusako A1 Kozuka-Hata, Hiroko A1 Oyama, Masaaki A1 Yoneda, Misako A1 Kai, ChiekoYR 2011 T1 Determination of a phosphorylation site in Nipah virus nucleoprotein and its involvement in virus transcription JF Journal of General Virology, VO 92 IS 9 SP 2133 OP 2141 DO https://doi.org/10.1099/vir.0.032342-0 PB Microbiology Society, SN 1465-2099, AB Many viruses use their host’s cellular machinery to regulate the functions of viral proteins. The phosphorylation of viral proteins is known to play a role in genome transcription and replication in paramyxoviruses. The paramyxovirus nucleoprotein (N), the most abundant protein in infected cells, is a component of the N–RNA complex and supports the transcription and replication of virus mRNA and genomic RNA. Recently, we reported that the phosphorylation of measles virus N is involved in the regulation of viral RNA synthesis. In this study, we report a rapid turnover of phosphorylation in the Nipah virus N (NiV-N). The phosphorylated NiV-N was hardly detectable in steady-state cells, but was detected after inhibition of cellular protein phosphatases. We identified a phosphorylated serine residue at Ser451 of NiV-N by peptide mass fingerprinting by electrospray ionization–quadrupole time-of-flight mass spectrometry. In the NiV minigenome assay, using luciferase as a reporter gene, the substitution of Ser451 for alanine in NiV-N resulted in a reduction in luciferase activity of approximately 45 % compared with the wild-type protein. Furthermore, the substitution of Ser451 for glutamic acid, which mimics a phosphoserine, led to a more significant decrease in luciferase activity – approximately 81 %. Northern blot analysis showed that both virus transcription and replication were reduced by these mutations. These results suggest that a rapid turnover of the phosphorylation of NiV-N plays an important role in virus transcription and replication., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/vir.0.032342-0