Project description:We report the genetic plasticity of Sendai virus and mumps virus. We introduced insertional mutation in the virus genome and checked fitness by comparing distribution of mutans in passage 1 and passage 2.
Project description:We evaluate the individual growth of 8 mutants of Sendai virus in the competitive condition under the cultured cells. We also evalate the growth of 6 mutants of mumps virus in the similar competitive condition.
Project description:Viral RNA synthesis of mononegaviruses occurs in the cytoplasmic membrane-less organelles called as inclusion body (IB). Here, we reported that the IBs of mumps virus (MuV), which is the causative agent of mumps and belongs to the family Paramyxoviridae, had properties of liquid organelles formed by liquid-liquid phase separation. Super-resolution microscopic analysis of the MuV IBs demonstrated that nucleocapsid and phospho-(P) proteins formed the cage-like structure and that viral polymerase was in a reticular pattern and partially co-localized with viral mRNA. In addition, we characterized host RNAs localized in the MuV IBs by a spatial transcriptome analysis and found that G-quadruplex motif sequences containing RNAs (G4-RNAs) were concentrated. In vitro phase separation assay showed that the G4-RNAs interacted with the P protein and enhanced condensation in the P droplets. Taken together, our data show that MuV generates the IBs with a characteristic cage-like structure and host G4-RNAs play an important role in the formation of MuV IBs
Project description:Mumps virus (MuV) is the etiological agent of mumps, a disease characterized by painful swelling of the parotid glands and often accompanied by severe complications. To understand the molecular mechanism of MuV infection, a functional analysis of the involved host factors is required. However, little is known about the host factors involved in MuV infection, especially those involved in the late stage of infection. In this project, we introduced the N-terminal and C-terminal fragments of split-TurboID into the cytoplasmic domains of hemagglutinin-neuraminidase and fusion proteins, respectively, and confirmed reconstitution and functional ligase activity of split-TurboID. Following affinity purification and mass spectrometry, 641 proteins were identified as being significantly enriched in the split-TurboID-expressing cells.
Project description:Recombinant Mumps viruses grown on Vero cells (Cercopithecus aethiops). Detection and quantification of defective viral genomes (DVGs) in high throughput sequencing data using DVG-profiler, a novel post sequence alignment processing algorithm.
