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Rabies virus M protein expressed inEscherichia coli and its regulatory role in virion-associated transcriptase activity

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Summary

Rabies virus M protein was expressed inEscherichia 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.

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Authors and Affiliations

  1. Department of Microbiology, Oita Medical University, Hasama-machi, Oita, Japan

    Y. Ito, A. Nishizono, K. Hiramatsu & K. Mifune

  2. Animal Laboratory Center, Oita Medical University, Hasama-machi, Oita, Japan

    K. Mannen

Authors
  1. Y. Ito
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  2. A. Nishizono
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  3. K. Mannen
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  4. K. Hiramatsu
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  5. K. Mifune
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Ito, Y., Nishizono, A., Mannen, K. et al. Rabies virus M protein expressed inEscherichia coli and its regulatory role in virion-associated transcriptase activity. Archives of Virology 141, 671–683 (1996). https://doi.org/10.1007/BF01718325

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  • DOI: https://doi.org/10.1007/BF01718325

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