Rabies virus M protein expressed in Escherichia coli and its regulatory role in virion-associated transcriptase activity.
Rabies virus M protein was expressed in Escherichia coli in the form of a fusion protein with maltose binding protein (MBP) and purified by amylose affinity column chromatography after extraction. In order to investigate the possible regulatory role of M protein in viral transcription, an assay system for rabies virion-associated transcriptase activity was established by using the ribonucleoprotein (RNP) cores prepared from purified virions. Analysis of the products of the transcription assay system showed that the products are sensitive to RNase and are positive-strand RNA. Addition of the fusion protein to the system after cleavage with a proteinase Factor Xa (FXa), which cleaves the fusion protein into the M protein and MBP, resulted in an efficient and dose-dependent inhibition of the transcription. Furthermore, addition to the system of anti-M protein monoclonal antibody significantly restored the transcription. Control experiments with the same transcription assaying system using rabies virus nucleoprotein expressed as a fusion protein with MBP and cleaved with FXa did not result in an inhibition of the transcription. These results suggest that the M protein of rabies virus has the property to down-regulate virion-associated transcription.
Ito
Y
Y
Department of Microbiology, Oita Medical University, Japan.
Nishizono
A
A
Mannen
K
K
Hiramatsu
K
K
Mifune
K
K
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D015536:Down-Regulation; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D015967:Gene Expression Regulation, Viral; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D011820:Rabies virus / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014158:Transcription, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000201:enzymology
D018528:ATP-Binding Cassette Transporters; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C461599:maltose transport system, E coli; D012321:DNA-Directed RNA Polymerases