Characterization of human parainfluenza virus type 2 RNAs in infected cells and by in vitro synthesis.
The RNA species synthesized in HPIV-2 infected CV-1 cells were identified and characterized. The largest RNA of approximately 5.5 x 10(6) in molecular weight (MW) based on electrophoretic mobility, was identified as the genomic RNA. The other small RNA species of MWs 2.4 x 10(6), 1.1 x 10(6), 0.77 x 10(6), 0.68 x 10(6) and 0.5 x 10(6) were identified as mRNAs. The five smallest RNAs were also synthesized in vitro and comigrated with RNAs synthesized in virus-infected cells. mRNAs synthesized both in vitro and in virus-infected cells were translated in vitro. NP, P, M and V proteins synthesized in vitro comigrated, when analyzed by SDS-PAGE, with the authentic proteins synthesized in virus-infected cells. Additionally, peptide mapping showed that the NP, P and M proteins synthesized in vitro were indistinguishable from their counterparts synthesized in infected cells. Analysis of the proteins from virions identified L, HN, NP, F (F1, F2), P, M and V proteins as virion structural proteins. Electrophoretic mobility of reduced and nonreduced F proteins was found to be different due to the conformational changes conferred by disulfide bonds.
Huang
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YT
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106-4907, USA.
Romito
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RR
Panin
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M
D002460:Cell Line; D006801:Humans; D066298:In Vitro Techniques; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis*; D018108:Respirovirus / Q000235:genetics*; D013696:Temperature; D013997:Time Factors; D015678:Viral Structural Proteins / Q000235:genetics
D012333:RNA, Messenger; D012367:RNA, Viral; D015678:Viral Structural Proteins