Intracellular cleavage of sigmaA protein of avian reovirus.
By Western blot analyzes of expression of avian reovirus proteins, one unknown fragment was detected by an anti-sigmaA monoclonal antibody in virus-infected cells lysate. It was interesting to note that RNA interference against sigmaA resulted in the suppression of the unknown fragment. Using various lengths of sigmaA constructs conjugated with different tags, we present evidences to demonstrate that the fragment comes from the cleavage of sigmaA and is the larger carboxyl-terminus, termed sigmaAC. Cleavage of sigmaA simultaneously produces a smaller amino-terminus, named sigmaAN. sigmaAC could be seen early in viral infection and accumulated with time and dose of infection, indicating that the derived products are not just transient intermediates of protein degradation. The same type of cleaved products were also observed in different genotypes and serotypes of ARV as well as in different cell lines, suggesting that this intracellular modification of sigmaA is common to all ARVs. Similar localization of sigmaAC in both cytosol and nucleus with sigmaA suggested that further modification of sigmaA may be important for its function. Our evidences suggest that besides the outer capsid protein muB, sigmaA may also have post-translational cleavage which has never been reported before even in related mammalian reovirus.
Ji
Wen T
WT
Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 912, Taiwan.
Lin
Feng L
FL
Wang
Ying C
YC
Shih
Wing L
WL
Lee
Long H
LH
Liu
Hung J
HJ
D000818:Animals; D001717:Birds; D015153:Blotting, Western; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D054884:Host-Pathogen Interactions; D030101:Orthoreovirus, Avian / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014758:Viral Core Proteins / Q000378:metabolism*
D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; C403292:sigmaA protein, avian reovirus