Immunoanalysis of antiviral Mx protein expression in Japanese flounder (Paralichthys olivaceus) cells.
Mx proteins are interferon-inducible GTPases that possess antiviral properties in vertebrates. Japanese flounder (Paralichthys olivaceus) Mx protein has previously been shown to possess some antiviral activity against rhabdoviruses. A polyclonal antibody was generated against a purified peptide fragment of Japanese flounder Mx protein that had been produced in an Escherichia coli expression system. The PAb detected the approximately 71 kDa Mx protein from Japanese flounder (hirame) natural embryo (HINAE) cells that had been cultured with poly I:C, an interferon inducer, but not in unstimulated cells. The polyclonal antibody did not cross react with Mx protein from carp epithelial, grouper fin and zebrafish embryo cell lines that had been similarly induced or transfected with poly I:C. By immunofluorescence cytochemistry, Japanese flounder Mx protein was localized to the cell cytoplasm. Hirame rhabdovirus stimulated expression of Mx protein in the infected and surrounding HINAE cells. Within virus-infected cells, there was some indication of Mx protein colocalizing with viral proteins. Poly I:C stimulation of HINAE cells induced an early increase in Mx protein mRNA transcripts, but maximum Mx mRNA transcript and protein expression was reached after 48 h. Both Mx mRNA transcripts and protein levels were maintained till at least 72 h.
Lin
Ooi Ei
OE
Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
Ohira
Tsuyoshi
T
Hirono
Ikuo
I
Saito-Taki
Tatsuo
T
Aoki
Takashi
T
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D005432:Flounder / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D019204:GTP-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D029141:Novirhabdovirus / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
D064547:Myxovirus Resistance Proteins; D012333:RNA, Messenger; D019204:GTP-Binding Proteins