Characterization of the GXXXG motif in the first transmembrane segment of Japanese encephalitis virus precursor membrane (prM) protein.
The interaction between prM and E proteins in flavivirus-infected cells is a major driving force for the assembly of flavivirus particles. We used site-directed mutagenesis to study the potential role of the transmembrane domains of the prM proteins of Japanese encephalitis virus (JEV) in prM-E heterodimerization as well as subviral particle formation. Alanine insertion scanning mutagenesis within the GXXXG motif in the first transmembrane segment of JEV prM protein affected the prM-E heterodimerization; its specificity was confirmed by replacing the two glycines of the GXXXG motif with alanine, leucine and valine. The GXXXG motif was found to be conserved in the JEV serocomplex viruses but not other flavivirus groups. These mutants with alanine inserted in the two prM transmembrane segments all impaired subviral particle formation in cell cultures. The prM transmembrane domains of JEV may play importation roles in prM-E heterodimerization and viral particle assembly.
Lin
Ying-Ju
YJ
Institute of Biotechnology, Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan. scwu@life.nthu.edu.tw
Peng
Jia-Guan
JG
Wu
Suh-Chin
SC
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D019281:Dimerization; D004664:Encephalitis Virus, Japanese / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D005416:Flavivirus / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics; D019065:Virus Assembly
D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus