Molecular structure and early events in the replication of Tacaribe arenavirus S RNA

Franze-Fernandez, Maria-T.; Zetina, Carlos R.; Iapalucci, Silvia; Lucero, Miguel A.; Bouissou, C.; López, Ricardo; Rey, Osvaldo; Daheli, Marcelo; Cohen, Georges N.; Zakin, Mario M.

doi:10.1016/0168-1702(87)90045-1

Cited by 85 publications

(64 citation statements)

References 29 publications

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“…Several studies have shown higher levels of NP mRNA than GPC mRNA throughout all times examined in LCMV-infected cells (27,28). Accordingly, CAT and GFP expression levels were higher when placed in lieu of NP than GPC (Fig.…”

Section: Bhk-21 Cells (Ifn -mentioning

confidence: 80%

Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

Emonet

Garidou

McGavern

et al. 2009

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

124

216

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Several arenaviruses cause hemorrhagic fever disease in humans for which no licensed vaccines are available and current therapeutic intervention is limited to the off-label use of the wide-spectrum antiviral ribavirin. However, the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) has proven to be a Rosetta stone for the investigation of virus-host interactions. Arenaviruses have a bisegmented negative-strand RNA genome. The S segment encodes for the virus nucleoprotein and glycoprotein, whereas the L segment encodes for the virus polymerase (L) and Z protein. The ability to generate recombinant LCMV (rLCMV) expressing additional foreign genes of interest would open novel avenues for the study of virushost interactions and the development of novel vaccine strategies and high-throughput screens to identify antiarenaviral molecules. To this end, we have developed a trisegmented (1L ؉ 2S) rLCMV-based approach (r3LCMV). Each of the two S segments in r3LCMV was altered to replace one of the viral genes by a gene of interest. All r3LCMVs examined expressing different reported genes were stable both genetically and phenotypically and exhibited wild-type growth properties in cultured cells. Reporter gene expression in r3LCMV-infected cells provided an accurate surrogate of levels of virus multiplication. Notably, some r3LCMVs displayed highly attenuated virulence in mice but induced protective immunity against a subsequent lethal challenge with wild-type LCMV, supporting the potential development of r3LCMV-based vaccines.antiviral screen ͉ reverse genetic ͉ viral attenuation A renaviruses merit significant interest both as tractable experimental model systems to study acute and persistent viral infections and as clinically important human pathogens (1-3). Thus, the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) has proven to be a superb workhorse in the field of virology and immunology (1). However, several arenaviruses cause hemorrhagic fever (HF) disease in humans, associated with high morbidity and mortality (2, 3). The Old World arenavirus Lassa virus (LASV) poses the highest public health concern among HF arenaviruses. LASV is estimated to infect several hundred thousand individuals yearly in its endemic region of West Africa, resulting in a high number of Lassa fever disease cases (2). Likewise, several New World arenaviruses, chiefly Junin virus, cause viral HF disease (3). Moreover, evidence indicates that the worldwide distributed LCMV is a neglected human pathogen of clinical significance (4) and poses a special threat to immunocompromised individuals (5, 6).Public health concerns posed by human pathogenic arenaviruses are aggravated by the lack of licensed vaccines and current therapy being limited to the use of the nucleoside analog ribavirin (Rib) that can cause significant side effects and requires an early and i.v. administration for optimal efficacy (3). Therefore, it is important to develop novel effective antiarenaviral drugs and vaccines, tasks that would be facilitated ...

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Section: Bhk-21 Cells (Ifn -mentioning

confidence: 80%

Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

Emonet

Garidou

McGavern

et al. 2009

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

124

216

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“…However, the GPs of LCMV and Lassa virus both are cleaved by the subtilase SKI-1/S1P. (24). The arrow indicates the cleavage site, which has been confirmed for Lassa virus GP-C (30), LCMV GP-C (ARM53b), Pichinde virus GP-C, and Tacaribe virus GP-C (13).…”

Section: Discussionmentioning

confidence: 99%

Endoproteolytic Processing of the Lymphocytic Choriomeningitis Virus Glycoprotein by the Subtilase SKI-1/S1P

Beyer

Pöpplau

Garten

et al. 2003

J Virol

212

210

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The envelope glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) is posttranslationally cleaved into two subunits. We show here that this endoproteolytic processing is not required for transport to the cell surface but is essential for LCMV GP to mediate infectivity of pseudotyped retroviral vectors. By systematic mutational analysis of the LCMV GP cleavage site, we determined that the consensus motif R-(R/K/H)-L-(A/L/S/T/F) 265 is essential for the endoproteolytic processing. In agreement with the identified consensus motif, we show that the cellular subtilase SKI-1/S1P cleaves LCMV GP.Lymphocytic choriomeningitis virus (LCMV) is the prototype of the Arenaviridae family, which includes important human pathogens causing hemorrhagic fever, such as Lassa virus, Junin virus, Machupo virus, and Guanarito virus. LCMV has been widely used as an experimental model for the study of immunology, viral persistence, and viral pathogenesis (34, 50).Virions of LCMV are composed of a nucleocapsid which is surrounded by a lipid envelope containing the envelope glycoprotein (GP). The initial steps in LCMV infection involve the interaction of GP with the cellular receptor of the target cells. Depending on the GP sequence, LCMV uses either alphadystroglycan or an alternative cellular protein as a receptor (15,41,42). After internalization of the virions within vesicles, LCMV GP mediates fusion of the viral and cellular membranes, resulting in delivery of the nucleocapsids into the cytoplasm (9,18,19).LCMV GP is initially expressed as a precursor polypeptide, GP-C, which is posttranslationally cleaved into two subunits, GP-1 and GP-2 (12). Cleavage of LCMV GP-C by a yetunidentified cellular protease occurs in the Golgi or a postGolgi compartment (47), whereas the related Lassa virus GP-C was found to be cleaved early in the secretory pathway by the subtilase SKI-1/S1P (4, 31). The amino-terminal cleavage product GP-1 is a peripheral membrane protein and is noncovalently associated with the carboxy-terminal subunit GP-2, which is an integral membrane protein (14). GP-1 presumably interacts with the cellular receptor, whereas the GP-2 subunit most likely mediates fusion of the viral envelope with the cellular membrane (8,13,26,27). The fusion peptide of GP-2 appears to be activated to a fusion-competent state by a pHdependent conformational change of the LCMV GP subunits (9,18,19).The GP of LCMV and other arenaviruses are cleaved between two uncharged amino acids (13, 30), whereas most other viral GPs are cleaved after a basic amino acid (29). In this study, we analyzed cleavage of LCMV GP in detail. We show that cleavage of LCMV GP is not required for transport to the cell surface but is required for mediating virus infectivity of retroviral pseudotypes. Furthermore, we have determined the consensus motif for cleavage of the LCMV GP, and we show that LCMV GP is cleaved in a late-Golgi or post-Golgi compartment by the cellular subtilase SKI-1/S1P. MATERIALS AND METHODS Construction of LCMV GP and Lassa virus GP expression ...

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“…By using cycloheximide or other protein synthesis inhibitors, it has been shown for several negativestrand RNA viruses that ongoing protein synthesis is required for replication of the viral genome, indicating that a virus-encoded protein is necessary for the synthesis of vc-and vRNA. Primary transcription of the infecting genome is, however, often not or only partly inhibited (Hay et al, 1977;Barrett et al, 1979;Abraham & Pattnaik, 1983;Pattnaik & Abraham, 1983;Patterson & Kolakofsky, 1984;Eshita et al, 1985;Ihara et al, 1985;Franze-Fernandez et al 1987;Shapiro et al, 1987). In the case of influenza virus, a member of the Orthomyxoviridae family, the N protein has been shown to anti-terminate transcription resulting in the synthesis of full-length vc-and vRNA, thus causing a switch from transcription to replication (Beaton & Krug, 1984, 1986.…”

Section: Discussionmentioning

confidence: 99%

Viral RNA synthesis in tomato spotted wilt virus-infected Nicotiana rustica plants

Kormelink¹,

Haan²,

Peters³

et al. 1992

Journal of General Virology

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The synthesis of viral RNA species in tomato spotted wilt virus-infected Nicotiana rustica plants was followed in terms of time and relative abundance. Systemic symptoms were visible after 4 days postinoculation (p.i.), but viral (v) and viral-complementary (vc) strands of all three genomic RNA segments [large (L) RNA, medium (M) RNA and small (S) RNA] were detected from 2 days p.i. In addition, two subgenomic mRNAs, derived from S RNA, were detected. For the L RNA segment no subgenomic mRNAs were detected, suggesting that this segment is expressed via the synthesis of a genome-sized vc mRNA. A possible M-specific subgenomic mRNA was detected, showing a similar time course of appearance as the subgenomic mRNAs derived from the S RNA segment. Analysis of cytoplasmic RNA fractions revealed that both v and vc strands of all three genomic segments associate with the nucleocapsid protein into nucleocapsid structures, the vcRNA species being present in lower amounts. Intact, enveloped virus particles contained only the v strand of the L RNA segment and, surprisingly, both v and vc strands of the M and S RNA segment, though in different ratios.

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Molecular structure and early events in the replication of Tacaribe arenavirus S RNA

Cited by 85 publications

References 29 publications

Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest

Endoproteolytic Processing of the Lymphocytic Choriomeningitis Virus Glycoprotein by the Subtilase SKI-1/S1P

Viral RNA synthesis in tomato spotted wilt virus-infected Nicotiana rustica plants

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