Reassortant Analysis of Guinea Pig Virulence of Pichinde Virus Variants

Zhang, Lihong; Marriott, Kathleen A.; Harnish, Delsworth G.; Aronson, Judith F.

doi:10.1006/viro.2001.1127

Cited by 34 publications

(36 citation statements)

References 23 publications

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“…A previous study [52] has shown that limited passages of viruses on cultured cells required for plaque purification did not change viral pathogenicity in vivo. We, therefore, believe that a single round of virus amplification should not greatly alter the native genomic sequences of the P2 and P18 viruses.…”

Section: Resultsmentioning

confidence: 93%

“…Published sequences of the S segment between the P2 and P18 strains have identified several sequence variations within the GPC and NP genes, suggesting that viral genomic sequence variations may dictate the differential behaviors of different strains of the virus in an infected host [51]. Reassortment experiments further suggest that genes encoded on both the S and L segments may contribute to the acquired virulence [52]. To our knowledge, the full-length sequences of the L segment for the P2 and P18 viruses are not yet available.…”

Section: Introductionmentioning

confidence: 99%

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Genome comparison of virulent and avirulent strains of the Pichinde arenavirus

et al. 2008

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A virulent (P18) strain of the Pichinde arenavirus produces a disease in guinea pigs that somewhat mimics human Lassa fever, whereas an avirulent (P2) strain of this virus is attenuated in infected animals. It has been speculated that the composition of viral genomes may confer the degree of virulence in an infected host; the complete sequence of the viral genomes, however, is not known. Here, we provide for the first time genomic sequences of both S and L segments for both the P2 and P18 strains. Sequence comparisons identify three mutations in the GP1 subunit of the viral glycoprotein, one in the nucleoprotein NP, and five in the viral RNA polymerase L protein. These mutations, alone or in combination, may contribute to the acquired virulence of Pichinde infection in animals. The 3 amino acid changes in the variable region of the GP1 glycoprotein subunit may affect viral entry by altering its receptor-binding activity. While NP has previously been shown to modulate the host immune responses to viral infection, we found that the R374K change in this protein does not affect the NP function in suppressing interferon-β expression. Four out of the five amino acid changes in the L protein occur in a small region of the protein that may contribute to viral virulence by enhancing its function on viral genomic RNA synthesis.

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Section: Resultsmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Genome comparison of virulent and avirulent strains of the Pichinde arenavirus

et al. 2008

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“…6): a hydrophilic N-terminal region comprising aa 1 to 39, followed by the 37-aa RING domain and a 19-aa C-terminal region. Comparison of the deduced protein sequence of all arenavirus Z genes sequenced thus far reveals a similar organization, with a conserved ring domain (51% homology) and less conserved N-and C-terminal regions (22 and 31% homology, respectively) (9,16,27,29). To assess whether the Z protein N-and C-terminal regions contribute to L binding and inhibition, we prepared Z mutants containing deletions at the C terminus, the N terminus, both the N and C termini, and the RING-plus-C-terminal regions.…”

Section: Tv Z Protein Interacts With L Proteinmentioning

confidence: 98%

“…Using this system, we also demonstrated that Z is a powerful inhibitor of RNA replication and transcription (21). Lymphocytic choriomeningitis virus (LCMV) Z protein exhibits a similar inhibitory effect when transcription and replication are analyzed in the homologous (LCMV) minigenome system (6).Arenavirus Z proteins contain a conserved 37-amino-acid (aa) RING domain that comprises about 40% of their amino acid residues (9,16,27,29). The RING domain binds two zinc ions in a cross-brace arrangement and is involved in proteinprotein interactions (2).…”

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confidence: 99%

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Tacaribe Virus Z Protein Interacts with the L Polymerase Protein To Inhibit Viral RNA Synthesis

Jácamo

López

Wilda

et al. 2003

J Virol

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Tacaribe virus (TV) is the prototype of the New World group of arenaviruses. The TV genome encodes four proteins, the nucleoprotein (N), the glycoprotein precursor, the polymerase (L), and a small RING finger protein (Z). Using a reverse genetic system, we recently demonstrated that TV N and L are sufficient to drive transcription and full-cycle RNA replication mediated by TV-like RNAs and that Z is a powerful inhibitor of these processes (N. López, R. Jácamo, and M. T. Franze-Fernández, J. Virol. 65:12241-12251, 2001). In the present study we investigated whether Z might interact with either of the proteins, N and L, required for RNA synthesis. To that end, we used coimmunoprecipitation with monospecific antibodies against the viral proteins and coimmunoprecipitation with serum against glutathione S-transferase (GST) and binding to glutathioneSepharose beads when Z was expressed as a fusion protein with GST. We demonstrated that Z interacted with L but not with N and that Z inhibitory activity was dependent on its ability to bind to L. We also evaluated the contribution of different Z regions to its binding ability and functional activity. We found that integrity of the RING structure is essential for Z binding to L and for Z inhibitory activity. Mutants with deletions at the N and C termini of Z showed that amino acids within the C-terminal region and immediately adjacent to the RING domain N terminus contribute to efficient Z-L interaction and are required for inhibitory activity. The data presented here provide the first evidence of an interaction between Z and L, suggesting that Z interferes with viral RNA synthesis by direct interaction with L. In addition, coimmunoprecipitation studies revealed a previously unreported interaction between N and L.Tacaribe virus (TV) is the prototype of the New World group of arenaviruses. Within this group the viruses form three phylogenetically distinct clades, one of which includes TV together with the four known South American pathogens that produce severe hemorrhagic disease (Junin, Machupo, Guanarito, and Sabia viruses) (5). TV, however, does not seem to be a human pathogen.TV, like all arenaviruses, is an enveloped virus containing two single-stranded RNA segments called S and L. The S RNA contains two genes, one encoding the nucleoprotein (N; 64 kDa) and one encoding the glycoprotein precursor (55 kDa) (10), while the L RNA encodes the RNA-dependent RNA polymerase (L; 240 kDa) (15) and a small protein with a RING finger motif (Z; 11 kDa) (16). In both S and L RNAs, the genes are arranged in opposite orientation and are separated by noncoding sequences that have the potential to form stable secondary structures (11). The 5Ј regions of arenavirus genomes and antigenomes, though positively stranded, are not translated directly into proteins. Rather, genomes and antigenomes are found only as nucleocapsids tightly bound to N, and the coding sequences are expressed from mRNAs transcribed from the 3Ј regions of the genomes or antigenomes (1,11,17,19,26). These mRNAs contai...

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