The large protein ``L' of Peste-des-petits-ruminants virus exhibits RNA triphosphatase activity, the first enzyme in mRNA capping pathway

Ansari, Mohammad Yunus and Singh, Piyush Kumar and Rajagopalan, Deepa and Shanmugam, Purnima and Bellur, Asutosh and Shaila, Melkote Subbarao (2019) The large protein ``L' of Peste-des-petits-ruminants virus exhibits RNA triphosphatase activity, the first enzyme in mRNA capping pathway. In: VIRUS GENES, 55 (1). pp. 68-75.

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Abstract

Peste-des-petits-ruminants is a highly contagious and fatal disease of goats and sheep caused by non-segmented, negative strand RNA virus belonging to the Morbillivirus genusPeste-des-petits-ruminants virus (PPRV) which is evolutionarily closely related to Rinderpest virus (RPV). The large protein L' of the members of this genus is a multifunctional catalytic protein, which transcribes and replicates the viral genomic RNA as well as possesses mRNA capping, methylation and polyadenylation activities; however, the detailed mechanism of mRNA capping by PPRV L protein has not been studied. We have found earlier that the L protein of RPV has RNA triphosphatase (RTPase), guanylyltransferase (GTase) and methyltransferase activities, and unlike vesicular stomatitis virus (VSV), follows the conventional pathway of mRNA capping. In the present work, using a 5-end labelled viral RNA as substrate, we demonstrate that PPRV L protein has RTPase activity when present in the ribonucleoprotein complex of purified virus as well as recombinant L-P complex expressed in insect cells. Further, a minimal domain in the C-terminal region (aa1640-1840) of the L protein has been expressed in E. coli and shown to exhibit RTPase activity. The RTPase activity of PPRV L protein is metal-dependent and functions with a divalent cation, either magnesium or manganese. In addition, RTPase associated nucleotide triphosphatase activity (NTPase) of PPRV L protein is also demonstrated. This work provides the first detailed study of RTPase activity and identifies the RTPase domain of PPRV L protein.

Item Type:	Journal Article
Publication:	VIRUS GENES
Publisher:	SPRINGER
Additional Information:	Copyright for this article belongs to Springer.
Keywords:	Morbillivirus; Peste-des-petits-ruminants virus L protein; PPRV; mRNA capping; RNA triphosphatase; Conventional mRNA capping
Department/Centre:	Division of Biological Sciences > Biochemistry Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	09 Mar 2019 09:11
Last Modified:	09 Mar 2019 09:11
URI:	http://eprints.iisc.ac.in/id/eprint/61900

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