[Co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus can form heterodimers].
In order to investigate the characterization of in vitro co-expressed GP5 and M proteins of porcine reproductive and respiratory syndrome virus (PRRSV), eukaryotic expression plasmids pCI-ORF5 (expressing GP5 protein alone), pCI-ORF6 (expressing M protein alone), and pCI-ORF5/ORF6 (co-expressing GP5 and M proteins) were constructed. After transient transfection, Western blot analysis under nonreducing condition demonstrated that co-expressed GP5 and M proteins could form disulfide-linked heterodimers (GP5-M) in transiently transfected BHK-21 cells. To further study the influence of GP5-M heterodimers formation on the subcellular localizations of GP5 or M proteins, green fluorescence protein (EGFP) and red fluorescence protein (RFP) were used as markers. The results of fluorescence distribution showed that co-expressed GP5-EGFP chimera and M-RFP chimera boosted the transport of GP5 from the endoplasmic reticulum (ER) to the Golgi complex, indicating that the formation of GP5-M heterodimers may be involved in posttranslational modification, transport, and subcellular localization of GP5. These results presented here lay foundation to further study the molecular mechanism of GP5-M heterodimer formation and its role in protective immunity of PRRSV.
Ma
Yan
Y
Laboratory of Animal Virology, College of Animal Science and Veterinary Medicine, China.
Jiang
Yun-bo
YB
Xiao
Shao-bo
SB
Fang
Liu-rong
LR
Chen
Huan-chun
HC
D000818:Animals; D015153:Blotting, Western; D019281:Dimerization; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013552:Swine; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*
C525999:M protein, porcine reproductive and respiratory syndrome virus; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; C114789:glycoprotein 5, PRRSV; D049452:Green Fluorescent Proteins