Characterization of infectious particles of grass carp reovirus by treatment with proteases.
Proteolytic cleavages play an important role in reovirus infection during entry into cells. The effects of protease digestion on the morphology, infectivity and polypeptide composition of grass carp reovirus (GCRV) were investigated. Following treatment with chymotrypsin, the different subviral particles of GCRV were isolated using density gradient centrifugation and examined by electron microscope (EM). Analysis of protein components revealed that the viral outer capsid was composed of VP5 and VP7. Of particular note, VP5 was found to primarily exist within virions as cleaved fragments, which was consistent with observations for its analogue mu1/mu1C, generated by autolysis of mu1 at the mu1N/mu1C junction for mammalian orthoreoviruses (MRVs). Meanwhile, both trypsin- and chymotrypsin-treated GCRV particles appeared to have an enhanced infectivity. Moreover, the corresponding assays between infectivity and protein component indicated that the enhancement of infectivity was correlated to the complete digestion of the outer capsid protein VP7 and partial cleavage of VP5. Overall, the results presented in this paper provided strong evidence that the proteins VP5 and VP7 of GCRV play an indispensable role in viral infection.
Fang
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Q
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China. qfang@wh.iov.cn
Seng
E K
EK
Ding
Q Q
QQ
Zhang
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LL
D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D002347:Carps / Q000821:virology; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002918:Chymotrypsin / Q000378:metabolism*; D007668:Kidney / Q000166:cytology; D046529:Microscopy, Electron, Transmission; D012087:Reoviridae / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D014357:Trypsin / Q000378:metabolism*; D014771:Virion / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure
D036022:Capsid Proteins; D002918:Chymotrypsin; D014357:Trypsin