Transfection-mediated generation of functionally competent Tula hantavirus with recombinant S RNA segment

Abstract

Since the discovery of RNA recombination in polioviruses, there has been a general belief that this mechanism operates only in positive-sense RNA viruses. Recently, studying wild-type Tula hantavirus, we observed a mosaic-like structure of the S RNA segment that was consistent with generation by recombination between viruses from two genetic lineages. Here we show transfection-mediated rescue of Tula virus carrying recombinant S RNA segment. Independent attempts yielded S RNA molecules of similar structure; the majority of them carried a break point located close to one of the break points suggested for natural recombinants. Recombinant virus puri®ed from the original variant was able to grow to the same titers in cell culture and showed the same characteristic immuno¯uorescence pattern when stained for the nucleocapsid protein. While competent, the recombinant virus appeared to be slightly less competitive than the wild type. Sequence analysis of the S cDNA clones obtained from the puri®ed recombinant virus con®rmed that all S RNA molecules were of recombinant origin. This provides the ®rst example of a negative-sense RNA virus constructed using homologous recombination.

Cytoplasmic tails of hantavirus glycoproteins interact with the nucleocapsid protein

“…The mapping results and previous findings lead us to propose a model in which both Gn and Gc, by their cytoplasmic tails, mediate binding to the RNP. Additionally, our data suggest that the N protein of PUUV interacts with Gn-and Gc-CTs of several hantavirus types, thus in theory enabling generation of reassortants via CT recognition of closely related viruses (Plyusnin et al, 2002;Razzauti et al, 2009). …”

“…Furthermore, the M segments have been shown to reassort upon cell-culture gene transfections (Plyusnin et al, 2002). Therefore, we studied whether the PUUV bacN and RNP are able to recognize Gn-CT peptides of different hantavirus strains.…”

Discovery of Severe Fever with Thrombocytopenia Syndrome Bunyavirus Strains Originating from Intragenic Recombination

“…However, intragenic recombination was considered being rare in negative-strand RNA virus (NSRV), including important human pathogens such as influenza, Ebola, and hantaviruses (4,10). These viruses have been thought to generate genetic diversity necessary for successful evolution, via mutation and reassortment of those with segmented genomes (19). Nevertheless, NSRV pathogens originating from HR and circulating in the field have been reported in influenza (12,13), Ebola (26), and hantaviruses (19,21).…”

“…These viruses have been thought to generate genetic diversity necessary for successful evolution, via mutation and reassortment of those with segmented genomes (19). Nevertheless, NSRV pathogens originating from HR and circulating in the field have been reported in influenza (12,13), Ebola (26), and hantaviruses (19,21). Coinfection experiments have also confirmed that HR can occur in the hantaviruses of the Bunyaviridae family (19).…”

“…Nevertheless, NSRV pathogens originating from HR and circulating in the field have been reported in influenza (12,13), Ebola (26), and hantaviruses (19,21). Coinfection experiments have also confirmed that HR can occur in the hantaviruses of the Bunyaviridae family (19). Here, our study suggests that HR results in the new SFTSV sublineage circulating in China, which further determines that HR is an important molecular mechanism of NSRV diversity.…”