The N-Terminal Domain of the Arenavirus L Protein Is an RNA Endonuclease Essential in mRNA Transcription

Morin, Benjamin; Coutard, Bruno; Lelke, Michaela; Ferrón, François; Kerber, Romy; Jamal, S.; Frangeul, Antoine; Baronti, Cécile; Charrel, Rémi N.; Lamballerie, Xavier de; Vonrhein, Clemens; Lescar, Julien; Bricogne, G.; Günther, Stephan; Canard, Bruno

doi:10.1371/journal.ppat.1001038

Cited by 158 publications

(184 citation statements)

References 51 publications

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“…The capped oligoribonucleotides are used as primers by the viral RdRp to initiate the transcription. The presence of an endonuclease domain in the RdRps of several other viruses, including SNV, has recently been proposed (47,61). However, it is not yet clear whether, similarly to that in influenza viruses, the entire cap-snatching process in these viruses is carried out solely by the RdRp or other host factors or whether viral proteins also play a role.…”

Section: Discussionmentioning

confidence: 99%

“…Unlike the situation in influenza virus, the RdRp of bunyaviruses and arenaviruses is encoded by one rather than three genes. Recent studies have suggested that RdRp from bunyaviruses and arenaviruses harbors the endonuclease domain at the N terminus, and its endonuclease activity has been demonstrated (47,61). Moreover, influenza viruses carry out cap snatching and transcription in the nucleus of infected cells, whereas bunyavirus and arenavirus transcription and genome replication are cytoplasmic.…”

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confidence: 99%

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Signatures of Host mRNA 5′ Terminus for Efficient Hantavirus Cap Snatching

Cheng

Mir

2012

J Virol

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Hantaviruses, similarly to other negative-strand segmented RNA viruses, initiate the synthesis of translation-competent capped mRNAs by a unique cap-snatching mechanism. Hantavirus nucleocapsid protein (N) binds to host mRNA caps and requires four nucleotides adjacent to the 5′ cap for high-affinity binding. N protects the 5′ caps of cellular transcripts from degradation by the cellular decapping machinery. The rescued 5′ capped mRNA fragments are stored in cellular P bodies by N, which are later efficiently used as primers by the hantaviral RNA-dependent RNA polymerase (RdRp) for transcription initiation. We showed that N also protects the host mRNA caps in P-body-deficient cells. However, the rescued caps were not effectively used by the hantavirus RdRp during transcription initiation, suggesting that caps stored in cellular P bodies by N are preferred for cap snatching. We examined the characteristics of the 5′ terminus of a capped test mRNA to delineate the minimum requirements for a capped transcript to serve as an efficient cap donor during hantavirus cap snatching. We showed that hantavirus RdRp preferentially snatches caps from the nonsense mRNAs compared to mRNAs engaged in translation. Hantavirus RdRp preferentially cleaves the cap donor mRNA at a G residue located 14 nucleotides downstream of the 5′ cap. The sequence complementarity between the 3′ terminus of viral genomic RNA and the nucleotides located in the vicinity of the cleavage site of the cap donor mRNA favors cap snatching. Our results show that hantavirus RdRp snatches caps from viral mRNAs. However, the negligible cap-donating efficiency of wild-type mRNAs in comparison to nonsense mRNAs suggests that viral mRNAs will not be efficiently used for cap snatching during viral infection due to their continuous engagement in protein synthesis. Our results suggest that efficiency of an mRNA to donate caps for viral mRNA synthesis is primarily regulated at the translational level.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Signatures of Host mRNA 5′ Terminus for Efficient Hantavirus Cap Snatching

Cheng

Mir

2012

J Virol

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“…The latter is a multidomain protein (5, 20) that harbors the viral RNA-dependent RNA polymerase (RdRp) (13, 17) and a cap-snatching endonuclease (22,29). It oligomerizes (34) and interacts with the viral promoter sequences (16,20).…”

Section: Lassa Virus (Lasv) Causing Hemorrhagic Lassa Fever In West Amentioning

confidence: 99%

Cross-Species Analysis of the Replication Complex of Old World Arenaviruses Reveals Two Nucleoprotein Sites Involved in L Protein Function

Kerber

Rieger

Busch

et al. 2011

J Virol

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Lassa virus (LASV) causing hemorrhagic Lassa fever in West Africa, Mopeia virus (MOPV) from EastAfrica, and lymphocytic choriomeningitis virus (LCMV) are the main representatives of the Old World arenaviruses. Little is known about how the components of the arenavirus replication machinery, i.e., the genome, nucleoprotein (NP), and L protein, interact. In addition, it is unknown whether these components can function across species boundaries. We established minireplicon systems for MOPV and LCMV in analogy to the existing LASV system and exchanged the components among the three systems. The functional and physical integrity of the resulting complexes was tested by reporter gene assay, Northern blotting, and coimmunoprecipitation studies. The family Arenaviridae comprises more than 20 virus species and is divided into the Old World and the New World complexes (9). Main representatives of the Old World complex are the prototype arenavirus lymphocytic choriomeningitis virus (LCMV), Lassa virus (LASV) causing hemorrhagic Lassa fever in West Africa (14), and Mopeia virus (MOPV) from East Africa. MOPV is closely related to LASV but, in contrast to the latter, is neither associated with human disease nor pathogenic in animal models (39).The genomes of arenaviruses consist of two negative-strand RNA segments. The large (L) segment and the small (S) segment each contain two genes in opposite orientation separated by an intergenic region (IGR). During genome replication, full-length copies of S and L RNA are synthesized. Viral transcripts contain a cap structure and terminate within the IGR (1). The IGR contains a stable secondary structure and plays a role in both transcription termination and virus assembly (31).The L RNA encodes the small Z protein and the 220-kDa L protein (26). The latter is a multidomain protein (5, 20) that harbors the viral RNA-dependent RNA polymerase (RdRp) (13, 17) and a cap-snatching endonuclease (22,29). It oligomerizes (34) and interacts with the viral promoter sequences (16,20). Z protein functions as a matrix protein (30, 37) and interacts with L protein (19,40). The S RNA encodes the glycoprotein precursor (GPC) and the nucleoprotein (NP). The virus genome, NP, and L protein represent the minimal cis-and trans-acting components required for replication and transcription (15,21,24). NP is the major component of the viral ribonucleoprotein (RNP) complex. It physically associates with itself as well as with L and Z proteins (7,19,23,35). Association with Z protein and with the ALIX/AIP1, an ESCRTassociated host protein, is important for incorporation of NP into virus-like particles (7,23,35,36). In addition, NP acts as an interferon antagonist (27,28). Recently published X-ray crystallographic data revealed a nucleotide binding site in the N terminus and a 3Ј-5Ј exoribonuclease in the C terminus of NP; the latter is critical for NP to function as an interferon antagonist (18,32).While a substantial amount of information has been accumulated on the individual components of the replication mac...

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“…Of these, influenza A virus, an orthomyxovirus, is the best studied and snatches the 59 end of pre-mRNAs in the nucleus (Herz et al 1981;Plotch et al 1981). Since bunyaviruses and arenaviruses replicate in the cytoplasm, they must use a distinct pool of mRNAs; however, while our mechanistic understanding of bunyaviral cap-snatching is increasing (van Poelwijk et al 1996;van Knippenberg et al 2005;Mir et al 2010;Morin et al 2010;Reguera et al 2010;Cheng and Mir 2012), little is known about whether the host can combat this replication step or what pool of endogenous mRNAs are being targeted for this process.…”

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confidence: 99%

A genome-wide RNAi screen reveals that mRNA decapping restricts bunyaviral replication by limiting the pools of Dcp2-accessible targets for cap-snatching

et al. 2013

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Bunyaviruses are an emerging group of medically important viruses, many of which are transmitted from insects to mammals. To identify host factors that impact infection, we performed a genome-wide RNAi screen in Drosophila and identified 131 genes that impacted infection of the mosquito-transmitted bunyavirus Rift Valley fever virus (RVFV). Dcp2, the catalytic component of the mRNA decapping machinery, and two decapping activators, DDX6 and LSM7, were antiviral against disparate bunyaviruses in both insect cells and adult flies. Bunyaviruses 59 cap their mRNAs by ''cap-snatching'' the 59 ends of poorly defined host mRNAs. We found that RVFV cap-snatches the 59 ends of Dcp2 targeted mRNAs, including cell cycle-related genes. Loss of Dcp2 allows increased viral transcription without impacting viral mRNA stability, while ectopic expression of Dcp2 impedes viral transcription. Furthermore, arresting cells in late S/early G2 led to increased Dcp2 mRNA targets and increased RVFV replication. Therefore, RVFV competes for the Dcp2-accessible mRNA pool, which is dynamically regulated and can present a bottleneck for viral replication.

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The N-Terminal Domain of the Arenavirus L Protein Is an RNA Endonuclease Essential in mRNA Transcription

Cited by 158 publications

References 51 publications

Signatures of Host mRNA 5′ Terminus for Efficient Hantavirus Cap Snatching

Signatures of Host mRNA 5′ Terminus for Efficient Hantavirus Cap Snatching

Cross-Species Analysis of the Replication Complex of Old World Arenaviruses Reveals Two Nucleoprotein Sites Involved in L Protein Function

A genome-wide RNAi screen reveals that mRNA decapping restricts bunyaviral replication by limiting the pools of Dcp2-accessible targets for cap-snatching

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