Complete sequence of the genome of the human isolate of Andes virus CHI-7913: comparative sequence and protein structure analysis

Tischler, Nicole D.; Fernández, Jorge Onrubia; Müller, Ilse; Martínez, Rodrigo; Galeno, H.; Villagra, Eliecer; Mora, Judith; Ramı́rez, Eugenio; Rosemblatt, Mario; Valenzuela, Pablo

doi:10.4067/s0716-97602003000200010

Cited by 17 publications

(12 citation statements)

References 29 publications

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“…The GPC-coding region of the cDNA of ANDV M segment strain CHI-7913 (Tischler et al, 2003) was cloned into the mammalian expression plasmid pI.18 (kindly provided by Jim Robertson from the National Institute for Biological Standards and Control, Hertfordshire, UK) as described previously (CifuentesMuñoz et al, 2010). Site-directed mutagenesis was performed using the QuikChange XL site-directed mutagenesis kit (Stratagene) as recommended by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

Aromatic and polar residues spanning the candidate fusion peptide of the Andes virus Gc protein are essential for membrane fusion and infection

Cifuentes-Muñoz

Barriga

Valenzuela

et al. 2010

Journal of General Virology

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Hantaviruses infect human cells through cell attachment and subsequent fusion of viral and cellular membranes at low pH. This largely unknown entry process is mediated by the Gn and Gc glycoproteins, anchored at the viral envelope membrane. Performing bioinformatic analysis and peptide-liposome-binding assays we suggested in a former report that Gc of Andes virus (ANDV) and other hantaviruses corresponds to the viral fusion protein sharing characteristics with class II fusion proteins. To gain insights into the fusion protein of hantaviruses, residues within the previously predicted fusion peptide of ANDV Gc were substituted and mutant proteins tested in fusion and infection assays. To ensure proper folding of mutant proteins, they were first characterized for trafficking to the plasma membrane and incorporation on to ANDV Gn/Gc-pseudotyped lentiviral particles. Cell attachment of these particles was assessed using a newly developed binding assay and their subsequent entry properties determined by FACS analysis of transduced cells expressing the GFP reporter gene. Furthermore, a three-colour-based cell-cell fusion assay of ANDV Gn/Gc expressing cells was performed. The results indicate an essential role of conserved Gc residues W115 and N118 in membrane fusion. Conversely, substitutions of the non-conserved Gc residue G116 did not considerably affect fusion and infection. Altogether, the findings are fully consistent with our earlier prediction suggesting Gc residues 115-121 as an internal fusion peptide and further emphasize the importance of aromatic and polar residues in hantavirus-cell membrane fusion.

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Section: Methodsmentioning

confidence: 99%

Aromatic and polar residues spanning the candidate fusion peptide of the Andes virus Gc protein are essential for membrane fusion and infection

Cifuentes-Muñoz

Barriga

Valenzuela

et al. 2010

Journal of General Virology

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“…The coding regions for the ANDV Gc glycoprotein amino (N)-and carboxyl (C)-terminal fragment were recovered from the cDNA clone of ANDV strain CHI-7913 (GenBank accession no. AY228238) by PCR amplification using primers 5Ј-GAAGGTCGACAAATGGCCGCAAGTGCAGAGACTCCA-3Ј and 5Ј-CA GTCTCGACCTAGCACCATTGTTTAAGGATGAC-3Ј to recover the N-terminal coding region (amino acids 647 to 866; GcA) and primers 5Ј-ATCCGTC GACTAATGCCATCTGTGAAGGTTTGTATA-3Ј and 5Ј-AGGACTCGAGG CGGCCGCTTAGACAGTTTTCTTGTGCCC-3Ј for the C-terminal region (amino acids 829 to 1138; GcB) (49). Escherichia coli BL21 cells were then transformed with plasmids pET32(a)/GcA and pET32(a)/GcB, which were generated by cloning of the PCR amplification products GcA and GcB into vector pET32(a) (Novagen; Merck Biosciences).…”

Section: Vol 83 2009 Andes Virus Antigens In Urine 5047mentioning

confidence: 99%

Andes Virus Antigens Are Shed in Urine of Patients with Acute Hantavirus Cardiopulmonary Syndrome

Godoy

Marsac

Stefas³

et al. 2009

J Virol

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Hantavirus cardiopulmonary syndrome (HCPS) is a highly pathogenic emerging disease (40% case fatality rate) caused by New World hantaviruses. Hantavirus infections are transmitted to humans mainly by inhalation of virus-contaminated aerosol particles of rodent excreta and secretions. At present, there are no antiviral drugs or immunotherapeutic agents available for the treatment of hantaviral infection, and the survival rates for infected patients hinge largely on early virus recognition and hospital admission and aggressive pulmonary and hemodynamic support. In this study, we show that Andes virus (ANDV) interacts with human apolipoprotein H (ApoH) and that ApoH-coated magnetic beads or ApoH-coated enzyme-linked immunosorbent assay plates can be used to capture and concentrate the virus from complex biological mixtures, such as serum and urine, allowing it to be detected by both immunological and molecular approaches. In addition, we report that ANDV-antigens and infectious virus are shed in urine of HCPS patients.

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“…The ARAUV Gn glycoprotein extended from aa 1 to 651 (nt 52-2004), including the conserved putative cleavage WAASA motif (aa 647-651), and the Gc glycoprotein extended from aa 652 to 1139 (nt 2005-3466). Five putative N-linked glycosylation sites at residues 138, 350, 402, 524 and 930 were predicted for ARAUV glycoprotein precursor as well as the three main O- Padula et al (2007) glycosylation clusters at residues 96, 306 and 582 (Tischler et al, 2003).…”

mentioning

confidence: 99%

Phylogenetic characterization of hantaviruses from wild rodents and hantavirus pulmonary syndrome cases in the state of Parana (southern Brazil)

Raboni

Hoffmann

Oliveira

et al. 2009

Journal of General Virology

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Over 1100 cases of hantavirus pulmonary syndrome (HPS) have occurred in Brazil since 1993, but little is known about Brazilian hantaviruses, and many of their rodent hosts remain unknown. The Araucaria hantavirus (ARAUV) was described recently from HPS patients from Paraná , in southern Brazil, but its host could not be identified. In this study, rodents were captured from regions with high HPS prevalence to address this issue. ARAUV RNA was detected in three distantly related rodent species: Oligoryzomys nigripes, Oxymycterus judex and Akodon montensis. Furthermore, a specimen of A. montensis was infected with a Jaborá -like virus, implying that A. montensis can be infected by at least two different hantaviruses. The presence of the same hantavirus strain in three different rodent species and the co-circulation of two different strains in the same rodent species highlight the potential for genomic reassortment, which could have an impact on hantavirus transmission dynamics in nature and on human epidemiology.Hantaviruses belong to the genus Hantavirus in the family Bunyaviridae and are found throughout most of the world. Similar to other members of the family, hantaviruses are enveloped viruses with an RNA genome comprised of three negative-sense, single-stranded segments. The large (L) RNA segment encodes an RNA-dependent RNA polymerase, the medium (M) segment encodes two envelope glycoproteins -Gn and Gc -processed from one precursor, and the small (S) segment encodes the nucleocapsid protein (N) (Plyusnin et al., 1996).Hantaviruses use small mammals as vectors, and in the wild, most of the different hantaviruses have been found to associate predominantly with a specific rodent species that acts as the host in a given geographical region (Plyusnin & Morzunov, 2001). Although sporadic spillover between rodents species has been suggested (Childs et al., 1994;Delfraro et al., 2008;Sousa et al.,. 2008), conclusive evidence that the studied viruses can establish productive infections in more than one rodent host species is still missing.Hantaviruses are transmitted to humans mainly through the inhalation of contaminated aerosols of rodent excreta (Lednicky, 2003;Schmaljohn & Patterson, 2001), but human-to-human transmission has also been described (Padula et al., 1998). These viruses are associated with two clinical syndromes, haemorrhagic fever with renal syndrome, described in Europe and Asia with a mortality rate of 1-15 %, and hantavirus pulmonary syndrome (HPS), described in the Americas with a mortality rate ranging from 40 to 60 % (Lundkvist & Niklasson, 1994;Johnson et al., 1999).Despite the increasing incidence of HPS in Brazil (Elkhoury, 2005), little is known about the genetic diversity of its causative agents, Brazilian hantaviruses (Table 1), and identification of their rodent hosts remains incomplete. The south-western region of the state of Paraná (southern Brazil, 26u 079 S 51u 319 W) is one of the most heavily affected areas of Brazil, with a high number of HPS cases: 168 cases have been reported ...

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Complete sequence of the genome of the human isolate of Andes virus CHI-7913: comparative sequence and protein structure analysis

Cited by 17 publications

References 29 publications

Aromatic and polar residues spanning the candidate fusion peptide of the Andes virus Gc protein are essential for membrane fusion and infection

Aromatic and polar residues spanning the candidate fusion peptide of the Andes virus Gc protein are essential for membrane fusion and infection

Andes Virus Antigens Are Shed in Urine of Patients with Acute Hantavirus Cardiopulmonary Syndrome

Phylogenetic characterization of hantaviruses from wild rodents and hantavirus pulmonary syndrome cases in the state of Parana (southern Brazil)

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