Transcription of a recombinant bunyavirus RNA template by transiently expressed bunyavirus proteins

Virology
E F DunnR M Elliott

Abstract

We describe a convenient system for analyzing bunyavirus transcription using a recombinant RNA template derived from the plasmid pBUNSCAT, which comprises a negative-sense reporter gene (chloramphenicol acetyltransferase or CAT) flanked by the exact 5' and 3' untranslated regions of the Bunyamwera virus (BUN) S RNA segment. When cells which expressed bunyavirus proteins (either by recombinant vaccinia viruses or by the vaccinia virus-T7 system) were transfected with BUNSCAT RNA, CAT activity could be measured, indicating transcription of the negative-sense reporter RNA into mRNA. The system permits investigation of both the protein and RNA sequence requirements for transcription. Extensions of 2 bases at the 5' end or 11 or 35 bases at the 3' end of BUNSCAT RNA allowed transcription but a lower level than the wild-type template. Deletion of the 5 nucleotides at the 3' end of BUNSCAT RNA reduced CAT activity by > 99%. Investigation of the viral protein requirements of the system showed that only the bunyavirus L and N proteins were needed for CAT activity. The BUN L protein was also able to transcribe the reporter RNA in concert with the N proteins of closely related bunyaviruses such as Batai, Cache Valley, Maguari, Main Drain,...Continue Reading

Citations

Dec 29, 2010·Proceedings of the National Academy of Sciences of the United States of America·Kaori TerasakiShinji Makino
Aug 28, 2009·Journal of Virology·Saleh A Eifan, Richard M Elliott
Oct 5, 2007·Journal of Virology·Monica M Ogg, Jean L Patterson
Jul 13, 2012·Journal of Virology·Erdong Cheng, Mohammad A Mir
Feb 13, 2001·Journal of Virology·A AlfadhliE Barklis
Apr 30, 2003·Journal of Virology·Ramon FlickFredrik Elgh
Jul 16, 2004·Journal of Virology·M A Mir, A T Panganiban
May 14, 2005·Journal of Virology·Anice C LowenRichard M Elliott
Sep 29, 2005·Journal of Virology·Vincent H J LeonardRichard M Elliott
Feb 24, 2001·Proceedings of the National Academy of Sciences of the United States of America·A BridgenR M Elliott
Dec 24, 1996·Proceedings of the National Academy of Sciences of the United States of America·A Bridgen, R M Elliott
Jan 10, 2009·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Ye ZhangXue-Fan Bai
Oct 31, 2008·The EMBO Journal·Mohammad A Mir, Antonito T Panganiban
Feb 28, 2014·Rapid Communications in Mass Spectrometry : RCM·Dale A ShepherdAlison E Ashcroft
Oct 22, 2002·The Journal of General Virology·Gabriele NeumannYoshihiro Kawaoka
Apr 20, 2012·The Journal of General Virology·Gjon BlakqoriRichard M Elliott
Nov 30, 2004·Virology·Anice C LowenRichard M Elliott
Feb 25, 2005·FEMS Microbiology Letters·Pramila Walpita, Ramon Flick
Jul 1, 2016·Annual Review of Phytopathology·Andrew O Jackson, Zhenghe Li
Sep 10, 2014·Nature Reviews. Microbiology·Richard M Elliott

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