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EMBO J. 2012 Mar 7;31(5):1320-9. doi: 10.1038/emboj.2011.483. Epub 2012 Jan 13.

Mechanism of RNA synthesis initiation by the vesicular stomatitis virus polymerase.

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1
Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115, USA.

Abstract

The minimal RNA synthesis machinery of non-segmented negative-strand RNA viruses comprises a genomic RNA encased within a nucleocapsid protein (N-RNA), and associated with the RNA-dependent RNA polymerase (RdRP). The RdRP is contained within a viral large (L) protein, which associates with N-RNA through a phosphoprotein (P). Here, we define that vesicular stomatitis virus L initiates synthesis via a de-novo mechanism that does not require N or P, but depends on a high concentration of the first two nucleotides and specific template requirements. Purified L copies a template devoid of N, and P stimulates L initiation and processivity. Full processivity of the polymerase requires the template-associated N protein. This work provides new mechanistic insights into the workings of a minimal RNA synthesis machine shared by a broad group of important human, animal and plant pathogens, and defines a mechanism by which specific inhibitors of RNA synthesis function.

PMID:
22246179
PMCID:
PMC3297992
DOI:
10.1038/emboj.2011.483
[Indexed for MEDLINE]
Free PMC Article
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