Short Double-stranded RNAs with an Overhanging 5′ ppp-Nucleotide, as Found in Arenavirus Genomes, Act as RIG-I Decoys

Marq, Jean‐Baptiste; Hausmann, Stéphane; Veillard, Nicolas; Kolakofsky, Daniel; Garcin, Dominique

doi:10.1074/jbc.m110.186262

Cited by 68 publications

(78 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Arenavirus genome synthesis uses a prime-and-realign mechanism (36), which results in a nontemplated 5=pppG that overhangs a panhandle structure predicted by the 3=-5=-terminal complementarity exhibited by arenavirus genomes. Interestingly, this unpaired 5=pppG overhang together with some mismatches within the predicted panhandle was shown to minimize the predicted double-stranded-RNA (dsRNA)-mediated induction of IFN-I and act as a RIG-I decoy (37). Therefore, it is plausible that the newly detected L-segment-derived RNA species in rLCMV (IGR/S-S)-infected cells cannot generate a dsRNA structure similar to the one associated with arenavirus genome and antigenome RNA species, which may have facilitated induction of IFN-I in rLCMV(IGR/S-S)-infected cells.…”

Section: Discussionmentioning

confidence: 99%

General Molecular Strategy for Development of Arenavirus Live-Attenuated Vaccines

Iwasaki

Ngo

Cubitt

et al. 2015

J Virol

View full text Add to dashboard Cite

Hemorrhagic fever arenaviruses (HFA) pose important public health problems in regions where they are endemic. Thus, Lassa virus (LASV) infects several hundred thousand individuals yearly in West Africa, causing a large number of Lassa fever cases associated with high morbidity and mortality. Concerns about human-pathogenic arenaviruses are exacerbated because of the lack of FDA-licensed arenavirus vaccines and because current antiarenaviral therapy is limited to an off-label use of ribavirin that is only partially effective. The Mopeia virus (MOPV)/LASV reassortant (ML29) is a LASV candidate live-attenuated vaccine (LAV) that has shown promising results in animal models. Nevertheless, the mechanism of ML29 attenuation remains unknown, which raises concerns about the phenotypic stability of ML29 in response to additional mutations. Development of LAVs based on well-defined molecular mechanisms of attenuation will represent a major step in combatting HFA. We used the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) to develop a general molecular strategy for arenavirus attenuation. Our approach involved replacement of the noncoding intergenic region (IGR) of the L genome segment with the IGR of the S genome segment to generate a recombinant LCMV, rLCMV(IGR/S-S), that was highly attenuated in vivo but induced protection against a lethal challenge with wild-type LCMV. Attenuation of rLCMV(IGR/S-S) was associated with a stable reorganization of the control of viral gene expression. This strategy can facilitate the rapid development of LAVs with the antigenic composition of the parental HFA and a mechanism of attenuation that minimizes concerns about increased virulence that could be caused by genetic changes in the LAV. IMPORTANCEHemorrhagic fever arenaviruses (HFA) cause high morbidity and mortality, and pose important public health problems in the regions where they are endemic. Implementation of live-attenuated vaccines (LAV) will represent a major step in combatting HFA. Here we have used the prototypic arenavirus LCMV to document a general molecular strategy for arenavirus attenuation that can facilitate the rapid development of safe and effective, as well as stable, LAV to combat HFA. Hemorrhagic fever arenaviruses (HFA) pose important public health problems in the regions where they are endemic (1-3). Thus, Lassa virus (LASV) infects several hundred thousand individuals yearly in West Africa, resulting in a high number of Lassa fever (LF) cases associated with high morbidity and significant mortality (4). Notably, increased traveling has led to the importation of LF cases into metropolitan areas around the globe where the virus is not endemic (5, 6). Moreover, regions of LASV endemicity are expanding (4), and the association of the recently identified arenavirus Lujo with a recent outbreak of HF in southern Africa (7,8) has raised concerns about the emergence of novel HFA. In addition, evidence indicates that the worldwide-distributed prototypic arenavirus lymphocytic choriomeningitis vi...

show abstract

Section: Discussionmentioning

confidence: 99%

General Molecular Strategy for Development of Arenavirus Live-Attenuated Vaccines

Iwasaki

Ngo

Cubitt

et al. 2015

J Virol

View full text Add to dashboard Cite

show abstract

“…Recent studies (24,25) suggest that unpaired pppG and mismatched bases in the panhandle region might allow arenaviruses to escape detection by the innate immune response by preventing the activation of RIG-I, an important immune sensor of LCMV infection (38). The panhandle configuration of the Lujo virus termini and the nature of nucleotide mismatches appear to be unique (Fig.…”

Section: Discussionmentioning

confidence: 99%

Reverse Genetics Recovery of Lujo Virus and Role of Virus RNA Secondary Structures in Efficient Virus Growth

Bergeron

Chakrabarti

Bird

et al. 2012

J Virol

View full text Add to dashboard Cite

Arenaviruses are rodent-borne viruses with a bisegmented RNA genome. A genetically unique arenavirus, Lujo virus, was recently discovered as the causal agent of a nosocomial outbreak of acute febrile illness with hemorrhagic manifestations in Zambia and South Africa. The outbreak had a case fatality rate of 80%. A reverse genetics system to rescue infectious Lujo virus from cDNA was established to investigate the biological properties of this virus. Sequencing the genomic termini showed unique nucleotides at the 3′ terminus of the S segment promoter element. While developing this system, we discovered that reconstructing infectious Lujo virus using the previously reported L segment intergenic region (IGR), comprising the arenaviral transcription termination signal, yielded an attenuated Lujo virus. Resequencing revealed that the correct L segment IGR was 36 nucleotides longer, and incorporating it into the reconstructed Lujo virus restored the growth rate to that of the authentic clinical virus isolate. These additional nucleotides were predicted to more than double the free energy of the IGR main stem-loop structure. In addition, incorporating the newly determined L-IGR into a replicon reporter system enhanced the expression of a luciferase reporter L segment. Overall, these results imply that an extremely stable secondary structure within the L-IGR is critical for Lujo virus propagation and viral protein production. The technology for producing recombinant Lujo virus now provides a method to precisely investigate the molecular determinants of virulence of this newly identified pathogen.

show abstract

“…This panhandle does not induce IFN upon transfection and, moreover, appears to act as a RIG-I decoy, presumably by binding but not activating RIG-I (Marq et al 2011). Alternative processing strategies are used by other negative-strand RNA viruses to generate genomic RNAs that fail to activate RIG-I.…”

Section: Importance Of Blunt-ended 59ppp-bprnamentioning

confidence: 99%

A structure-based model of RIG-I activation

Kolakofsky

Kowalinski

Cusack

2012

RNA

Self Cite

139

134

View full text Add to dashboard Cite

show abstract

Short Double-stranded RNAs with an Overhanging 5′ ppp-Nucleotide, as Found in Arenavirus Genomes, Act as RIG-I Decoys

Cited by 68 publications

References 32 publications

General Molecular Strategy for Development of Arenavirus Live-Attenuated Vaccines

General Molecular Strategy for Development of Arenavirus Live-Attenuated Vaccines

Reverse Genetics Recovery of Lujo Virus and Role of Virus RNA Secondary Structures in Efficient Virus Growth

A structure-based model of RIG-I activation

Contact Info

Product

Resources

About