Novel strategies for development of hemorrhagic fever arenavirus live-attenuated vaccines

Martínez-Sobrido, Luis; Torre, Juan Carlos de la

doi:10.1080/14760584.2016.1182024

Cited by 5 publications

(2 citation statements)

References 60 publications

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“…Lymphocytic choriomeningitis virus (LCMV), a model for arenavirus biology and mapping of the cellular immune responses, is globally seroprevalent and responsible for febrile illness, aseptic meningitis, neonatal disease, and multiple organ failure in transplant patients ( 3 – 9 ). Although promising vaccine candidates ( 10 – 13 ) and therapeutic treatments ( 14 – 16 ) have been developed to combat arenavirus diseases, to date there are limited effective intervention strategies for infected individuals.…”

Section: Introductionmentioning

confidence: 99%

Isolation of Reconstructed Functional Ribonucleoprotein Complexes of Machupo Virus

Pyle

Whelan

2021

J Virol

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Arenaviruses initiate infection by delivering a transcriptionally-competent ribonucleoprotein (RNP) complex into the cytosol of host cells. The arenavirus RNP consists of the large (L) RNA-dependent RNA polymerase (RdRP) bound to a nucleoprotein (NP)-encapsidated genomic RNA (vRNA) template. During transcription and replication, L must transiently displace RNA-bound NP to allow for template access into the RdRP active site. Concomitant with RNA replication new subunits of NP must be added to the nascent complementary RNAs (cRNA) as they emerge from the product exit channel of L. Interactions between L and NP thus play a central role in arenavirus gene expression. We developed an approach to purify recombinant functional RNPs from mammalian cells in culture using a synthetic vRNA, affinity-tagged L and NP. Negative-stain electron microscopy of purified RNPs revealed they adopt diverse and flexible structures, like RNPs of other Bunyavirales members. Monodisperse L-NP and trimeric ring-like NP complexes were also obtained in excess of flexible RNPs, suggesting that these heterodimeric structures self-assemble in the absence of suitable RNA templates. This work allows for further biochemical analysis of the interaction between arenavirus L and NP proteins and provides a framework for future high-resolution structural analyses of this replication-associated complex. IMPORTANCE Arenaviruses are rodent-borne pathogens that can cause severe disease in humans. All arenaviruses begin the infection cycle with delivery of the virus replication machinery into the cytoplasm of the host cell. This machinery consists of an RNA-dependent RNA polymerase – which copies the viral genome segments and synthesizes all four viral mRNAs – bound to the two nucleoprotein-encapsidated genomic RNAs. How this complex assembles remains a mystery. Our findings provide direct evidence for the formation of diverse intracellular arenavirus replication complexes using purification strategies for the polymerase, nucleoprotein, and genomic RNA of Machupo virus, which causes Bolivian hemorrhagic fever in humans. We demonstrate that the polymerase and nucleoprotein assemble into higher-order structures within cells, providing a model for the molecular events of arenavirus RNA synthesis. These findings provide a framework for probing the architectures and functions of the arenavirus replication machinery, and thus advancing antiviral strategies targeting this essential complex.

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Section: Introductionmentioning

confidence: 99%

Isolation of Reconstructed Functional Ribonucleoprotein Complexes of Machupo Virus

Pyle

Whelan

2021

J Virol

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“…The L segment codes for the RNA-dependent RNA polymerase (L) and the small matrix protein (Z), while the S segment encodes the nucleocapsid protein (N) and a glycoprotein precursor (GPC). Some members of this family cause hemorrhagic fever disease in humans and represent an important public health concern in their endemic regions, such as Lassa and Lujo viruses in Africa, Sabiá virus in Brazil, Guanarito virus in Venezuela, Chaparé and Machupo viruses (MACV) in Bolivia, and Junín virus (JUNV) in Argentina [ 1 , 2 ]. JUNV is the etiological agent of Argentine hemorrhagic fever (AHF), with the administration of immune plasma being the only therapeutic intervention against the disease.…”

Section: Introductionmentioning

confidence: 99%

Role of the ERK1/2 Signaling Pathway in the Replication of Junín and Tacaribe Viruses

Brunetti

Foscaldi

Quintana

et al. 2018

Viruses

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We have previously shown that the infection of cell cultures with the arenaviruses Junín (JUNV), Tacaribe (TCRV), and Pichindé promotes the phosphorylation of mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinases 1 and 2 (ERK1/2) and that this activation is required for the achievement of a productive infection. Here we examined the contribution of ERK1/2 in early steps of JUNV and TCRV multiplication. JUNV adsorption, internalization, and uncoating were not affected by treatment of cultured cells with U0126, an inhibitor of the ERK1/2 signaling pathway. In contrast, U0126 caused a marked reduction in viral protein expression and RNA synthesis, while JUNV RNA synthesis was significantly augmented in the presence of an activator of the ERK1/2 pathway. Moreover, U0126 impaired the expression of a reporter gene in a TCRV-based replicon system, confirming the ability of the compound to hinder arenavirus macromolecular synthesis. By using a cell-based assay, we determined that the inhibitor did not affect the translation of a synthetic TCRV-like mRNA. No changes in the phosphorylation pattern of the translation factor eIF2α were found in U0126-treated cells. Our results indicate that U0126 impairs viral RNA synthesis, thereby leading to a subsequent reduction in viral protein expression. Thus, we conclude that ERK1/2 signaling activation is required for an efficient arenavirus RNA synthesis.

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Bacterial Artificial Chromosome Reverse Genetics Approaches for SARS-CoV-2

Chiem,

Nogales,

Almazán

et al. 2023

Methods in Molecular Biology

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Novel strategies for development of hemorrhagic fever arenavirus live-attenuated vaccines

Cited by 5 publications

References 60 publications

Isolation of Reconstructed Functional Ribonucleoprotein Complexes of Machupo Virus

Isolation of Reconstructed Functional Ribonucleoprotein Complexes of Machupo Virus

Role of the ERK1/2 Signaling Pathway in the Replication of Junín and Tacaribe Viruses

Bacterial Artificial Chromosome Reverse Genetics Approaches for SARS-CoV-2

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