Scientific Reports
| Autor: | Courtney P. Long, Leslie E. W. LaConte, Sarah M. McDonald, Shu Zhang, Rebecca M. Mingo |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Rotavirus
0301 basic medicine replication genotype viruses In silico DNA Mutational Analysis Reassortment lcsh:Medicine Genome Viral Viral Nonstructural Proteins system Biology Virus Replication Genome Article Cell Line reverse genetics 03 medical and health sciences Animals Humans rearrangements wa-like lcsh:Science Gene candidate Recombination Genetic 2. Zero hunger Genetics mechanisms Multidisciplinary lcsh:R RNA-Binding Proteins RNA Haplorhini Reverse Genetics Reverse genetics 3. Good health constellations 030104 developmental biology Viral replication Helper virus lcsh:Q protein Reassortant Viruses |
| Zdroj: | Scientific Reports, Vol 7, Iss 1, Pp 1-12 (2017) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Rotaviruses (RVs) can evolve through the process of reassortment, whereby the 11 double-stranded RNA genome segments are exchanged among strains during co-infection. However, reassortment is limited in cases where the genes or encoded proteins of co-infecting strains are functionally incompatible. In this study, we employed a helper virus-based reverse genetics system to identify NSP2 gene regions that correlate with restricted reassortment into simian RV strain SA11. We show that SA11 reassortants with NSP2 genes from human RV strains Wa or DS-1 were efficiently rescued and exhibit no detectable replication defects. However, we could not rescue an SA11 reassortant with a human RV strain AU-1 NSP2 gene, which differs from that of SA11 by 186 nucleotides ( 36 amino acids). To map restriction determinants, we engineered viruses to contain chimeric NSP2 genes in which specific regions of AU-1 sequence were substituted with SA11 sequence. We show that a region spanning AU-1 NSP2 gene nucleotides 784-820 is critical for the observed restriction; yet additional determinants reside in other gene regions. In silico and in vitro analyses were used to predict how the 784-820 region may impact NSP2 gene/protein function, thereby informing an understanding of the reassortment restriction mechanism. Virginia Tech Carilion Research Institute; National Institutes of Health [R01-AI116815, R21-AI119588]; Translational Biology, Medicine, and Health Graduate Program at Virginia Tech The authors would like to thank members of the McDonald laboratory for intellectual and technical support. We also thank Dr. John Patton (University of Maryland, College Park) for the generous donation of reagents. This work was supported through start-up funding from the Virginia Tech Carilion Research Institute and through grants from the National Institutes of Health (R01-AI116815 and R21-AI119588). C.P.L. was also supported by the Translational Biology, Medicine, and Health Graduate Program at Virginia Tech. |
| Databáze: | OpenAIRE |
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