Sendai Virus RNA-dependent RNA Polymerase L Protein Catalyzes Cap Methylation of Virus-specific mRNA

Ogino, Tomoaki; Kobayashi, Masaki; Iwama, Minako; Mizumoto, Kiyohisa

doi:10.1074/jbc.m411167200

Cited by 95 publications

(94 citation statements)

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“…RTPase activity of L protein, the first step in viral mRNA capping, was tested in vitro using synthetic c-32 P-labelled RNA substrate containing sequences corresponding to viral genes. Viral mRNA 59 cis sequences were used as the substrate because in many negative-sense RNA viruses, the capping machinery of these viruses requires specific cis-acting signals in the RNA (Ogino et al, 2005;Ogino & Banerjee, 2007;Wang et al, 2007). Under optimal conditions, L protein in the RNP complex, as well as L protein co-expressed with P protein in insect cells, efficiently catalyses the removal of cphosphate from the triphosphate-ended RNA (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…In negative-sense RNA viruses, L protein is believed to possess activities required for capping, methylation and polyadenylation of mRNAs (Banerjee, 1987). Purified ribonucleoprotein (RNP) complexes from vesicular stomatitis virus (VSV) and Sendai virus containing the N, L and P proteins along with the genomic RNA have been shown to catalyse capping as well as cap methylation activities in vitro (Galloway et al, 2008;Grdzelishvili et al, 2006;Li et al, 2006;Ogino et al, 2005;Ogino & Banerjee, 2007. However, there is no direct demonstration of RTPase or GT activity associated with the RNP complex or L protein of any paramyxovirus.…”

Section: Introductionmentioning

confidence: 99%

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RNA triphosphatase and guanylyl transferase activities are associated with the RNA polymerase protein L of rinderpest virus

Gopinath

Shaila

2009

Journal of General Virology

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Rinderpest virus (RPV) large (L) protein is an integral part of the ribonucleoprotein (RNP) complex of the virus that is responsible for transcription and replication of the genome. Previously, we have shown that recombinant L protein coexpressed along with P protein (as the L-P complex) catalyses the synthesis of all viral mRNAs in vitro and the abundance of mRNAs follows a gradient of polarity, similar to the occurrence in vivo. In the present work, we demonstrate that the viral mRNAs synthesized in vitro by the recombinant L or purified RNP are capped and methylated at the N 7 guanine position. RNP from the purified virions, as well as recombinant L protein, shows RNA triphosphatase (RTPase) and guanylyl transferase (GT) activities. L protein present in the RNP complex catalyses the removal of c-phosphate from triphosphate-ended 25 nt RNA generated in vitro representing the viral N-terminal mRNA 59 sequence. The L protein forms a covalent enzyme-guanylate intermediate with the GMP moiety of GTP, whose formation is inhibited by the addition of pyrophosphate; thus, it exhibits characteristics of cellular GTs. The covalent bond between the enzyme and nucleotide is acid labile and alkali stable, indicating the presence of phosphoamide linkage. The C-terminal region (aa 1717-2183) of RPV L protein alone exhibits the first step of GT activity needed to form a covalent complex with GMP, though it lacks the ability to transfer GMP to substrate RNA. Here, we describe the biochemical characterization of the newly found RTPase/GT activity of L protein.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

RNA triphosphatase and guanylyl transferase activities are associated with the RNA polymerase protein L of rinderpest virus

Gopinath

Shaila

2009

Journal of General Virology

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“…On the other hand, within the K1220, R1221, and R1233 residues, which are conserved only in the L proteins of vesiculoviruses (e.g., VSV, Chandipura virus), lyssaviruses (e.g., rabies virus), and ephemerovirus (bovine ephemeral fever virus) belonging to the Rhabdoviridae family (Fig. S5), only the R1221 residue was found to be important for the L-pRNA complex formation (lanes [16][17][18], suggesting that the R1221 residue is required for some step of the complex formation, e,g., recognition of the rhabdovirus-specific mRNA-start sequence (5′-AACA-). The H360 (in the GHP motif) and H639 [in the G(x)8H motif] residues that are conserved histidine residues located in the blocks I and III, respectively, were not required for the L-pRNA complex formation (lanes 21 and 22).…”

Section: The Hr Motif In the L Protein Is Required For The Prntase Acmentioning

confidence: 99%

Histidine-mediated RNA transfer to GDP for unique mRNA capping by vesicular stomatitis virus RNA polymerase

Ogino

Yadav

Banerjee

2010

Proc. Natl. Acad. Sci. U.S.A.

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201

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The RNA-dependent RNA polymerase L protein of vesicular stomatitis virus, a prototype of nonsegmented negative-strand (NNS) RNA viruses, forms a covalent complex with a 5′-phosphorylated viral mRNA-start sequence (L-pRNA), a putative intermediate in the unconventional mRNA capping reaction catalyzed by the RNA:GDP polyribonucleotidyltransferase (PRNTase) activity. Here, we directly demonstrate that the purified L-pRNA complex transfers pRNA to GDP to produce the capped RNA (Gpp-pRNA), indicating that the complex is a bona fide intermediate in the RNA transfer reaction. To locate the active site of the PRNTase domain in the L protein, the covalent RNA attachment site was mapped. We found that the 5′-monophosphate end of the RNA is linked to the histidine residue at position 1,227 (H1227) of the L protein through a phosphoamide bond. Interestingly, H1227 is part of the histidine-arginine (HR) motif, which is conserved within the L proteins of the NNS RNA viruses including rabies, measles, Ebola, and Borna disease viruses. Mutagenesis analyses revealed that the HR motif is required for the PRNTase activity at the step of the enzyme-pRNA intermediate formation. Thus, our findings suggest that an ancient NNS RNA viral polymerase has acquired the PRNTase domain independently of the eukaryotic mRNA capping enzyme during evolution and PRNTase becomes a rational target for designing antiviral agents.nonsegmented negative-strand RNA virus | polyribonucleotidyltransferase | RNA-dependent RNA polymerase | L protein | mRNA modification A structural hallmark of eukaryotic mRNA is the presence of the 5′-terminal cap structure ½m 7 Gð5 0 Þpppð5 0 ÞN-, in which 7-methylguanosine (m 7

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“…In comparison with HRSV, metapneumoviruses lack the non-structural proteins NS1 and NS2, and the order of genes between the matrix (M) and large polymerase (L) is different: 39-le-N-P-M-F-M2-SH-G-Ltr-59 for HMPV and AMPV, and 39-le-NS1-NS2-N-P-M-SH-G-F-M2-L-tr-59 for HRSV. (Grdzelishvili et al, 2005;Hercyk et al, 1988;Ogino et al, 2005). The assembly and polymerase co-factor P and the L protein form the minimal complex needed for viral polymerase activity (Mazumder & Barik, 1994).…”

Section: Introductionmentioning

confidence: 99%

“…This RNP contains the viral genomic RNA (vRNA) encapsidated by the nucleocapsid protein (N), the phosphoprotein (P) and the large polymerase protein (L). By analogy with other paramyxoviruses, the L protein is responsible for the main catalytic activities of the viral polymerase complex Grdzelishvili et al, 2005;Hercyk et al, 1988;Ogino et al, 2005). The assembly and polymerase co-factor P and the L protein form the minimal complex needed for viral polymerase activity (Mazumder & Barik, 1994).…”

mentioning

confidence: 99%

Specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses

Graaf

Herfst

Schrauwen

et al. 2008

Journal of General Virology

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Human metapneumovirus (HMPV) and avian metapneumovirus (AMPV) have a similar genome organization and protein composition, but a different host range. AMPV subgroup C (AMPV-C) is more closely related to HMPV than other AMPVs. To investigate the specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses, a minireplicon system was generated for AMPV-C and used in combination with minireplicon systems for HMPV lineages A1 and B1. Viral RNA-like molecules representing HMPV-A1 and -B1, AMPV-A and -C and human respiratory syncytial virus were replicated efficiently by polymerase complexes of HMPV-A1 and -B1 and AMPV-C, but not by polymerase complexes of bovine parainfluenza virus 3. Upon exchange of HMPV and AMPV-C polymerase complex components, all chimeric polymerase complexes were functional; exchange between HMPVs did not result in altered polymerase activity, whereas exchange between HMPVs and AMPV-C did. Recombinant HMPV-B1 viruses in which polymerase genes were exchanged with those of HMPV-A1 replicated with normal kinetics in vitro, whilst replacement with AMPV-C genes resulted in moderate differences in virus replication. In hamsters, recombinant HMPV-B1 viruses in which individual polymerase genes were exchanged with those of AMPV-C were attenuated, irrespective of the results obtained with minireplicon systems or in vitro replication assays. This study provides insight into the specificity and functional interaction of polymerase complex proteins of human and avian metapneumoviruses, but neither minireplicon systems nor in vitro replication kinetics were found to be predictive for attenuation in permissive animals. INTRODUCTIONHuman metapneumovirus (HMPV) is an enveloped, nonsegmented, negative-strand RNA virus that causes respiratory tract illnesses, primarily in infants, young children, the frail elderly, and immunocompromised individuals (Crowe, 2004;Falsey et al., 2003;Kahn, 2006;Pelletier et al., 2002;van den Hoogen et al., 2001 van den Hoogen et al., , 2003. HMPV is a member of the family Paramyxoviridae, subfamily Pneumovirinae, genus Metapneumovirus, and can be divided in two main genetic lineages (A and B) each consisting of two sublineages, A1 and A2, and B1 and B2 (van den Hoogen et al., 2004). The only other member of the genus Metapneumovirus is avian metapneumovirus (AMPV). AMPV has been found to infect domestic poultry worldwide, causing acute respiratory infections (Cook, 2000). AMPVs have been classified into four subgroups, A-D (Bayon-Auboyer et al., 1999;Eterradossi et al., 1995;Juhasz & Easton, 1994;Seal, 1998). AMPV-C was first detected in the USA and is more closely related to HMPV than the other AMPV subgroups (Govindarajan & Samal, 2004Govindarajan et al., 2004;Toquin et al., 2003;van den Hoogen et al., 2002;Yunus et al., 2003). Human respiratory syncytial virus (HRSV) is the only other member of the subfamily Pneumovirinae that infects humans. In comparison with HRSV, metapneumoviruses lack the non-structural proteins NS1 and NS2,...

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Sendai Virus RNA-dependent RNA Polymerase L Protein Catalyzes Cap Methylation of Virus-specific mRNA

Cited by 95 publications

References 41 publications

RNA triphosphatase and guanylyl transferase activities are associated with the RNA polymerase protein L of rinderpest virus

RNA triphosphatase and guanylyl transferase activities are associated with the RNA polymerase protein L of rinderpest virus

Histidine-mediated RNA transfer to GDP for unique mRNA capping by vesicular stomatitis virus RNA polymerase

Specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses

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