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

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.

• Authors: Benjamin Morin, Amal A Rahmeh, Sean P J Whelan
• Journal: The EMBO journal (EMBO J) Vol. 31 Issue 5 Pg. 1320-9 (Mar 07 2012) ISSN: 1460-2075 [Electronic] England
• PMID: 22246179 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Viral
• Recombinant Proteins
• Viral Proteins
• DNA-Directed RNA Polymerases

Topics

• DNA-Directed RNA Polymerases (isolation & purification, metabolism)
• Models, Biological
• RNA, Viral (metabolism)
• Recombinant Proteins (isolation & purification, metabolism)
• Vesiculovirus (enzymology)
• Viral Proteins (isolation & purification, metabolism)