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A Single Point Mutation, Asn₁₆?Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

ABSTRACT: Studies of conditionally lethal mutants can help delineate the structure-function relationships of biomolecules. Temperature-sensitive (ts) mammalian reovirus (MRV) mutants were isolated and characterized many years ago. Two of the most well-defined MRV ts mutants are tsC447, which contains mutations in the S2 gene encoding viral core protein ?2, and tsG453, which contains mutations in the S4 gene encoding major outer-capsid protein ?3. Because many MRV ts mutants, including both tsC447 and tsG453, encode multiple amino acid substitutions, the specific amino acid substitutions responsible for the ts phenotype are unknown. We used reverse genetics to recover recombinant reoviruses containing the single amino acid polymorphisms present in ts mutants tsC447 and tsG453 and assessed the recombinant viruses for temperature-sensitivity by efficiency-of-plating assays. Of the three amino acid substitutions in the tsG453 S4 gene, Asn₁₆-Lys was solely responsible for the tsG453ts phenotype. Additionally, the mutant tsC447 Ala₁₈₈-Val mutation did not induce a temperature-sensitive phenotype. This study is the first to employ reverse genetics to identify the dominant amino acid substitutions responsible for the tsC447 and tsG453 mutations and relate these substitutions to respective phenotypes. Further studies of other MRV ts mutants are warranted to define the sequence polymorphisms responsible for temperature sensitivity.

SUBMITTER: Glover KKM

PROVIDER: S-EPMC7917769 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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A Single Point Mutation, Asn16→Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

Glover Kathleen K M KKM Sutherland Danica M DM Dermody Terence S TS Coombs Kevin M KM

Viruses 20210212 2

Studies of conditionally lethal mutants can help delineate the structure-function relationships of biomolecules. Temperature-sensitive (ts) mammalian reovirus (MRV) mutants were isolated and characterized many years ago. Two of the most well-defined MRV ts mutants are tsC447, which contains mutations in the S2 gene encoding viral core protein σ2, and tsG453, which contains mutations in the S4 gene encoding major outer-capsid protein σ3. Because many MRV ts muta ...[more]

PMID: 33673179

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Project description:UnlabelledReceptors expressed on the host cell surface adhere viruses to target cells and serve as determinants of viral tropism. Several viruses bind cell surface glycans to facilitate entry, but the contribution of specific glycan moieties to viral disease is incompletely understood. Reovirus provides a tractable experimental model for studies of viral neuropathogenesis. In newborn mice, serotype 1 (T1) reovirus causes hydrocephalus, whereas serotype 3 (T3) reovirus causes encephalitis. T1 and T3 reoviruses engage distinct glycans, suggesting that glycan-binding capacity contributes to these differences in pathogenesis. Using structure-guided mutagenesis, we engineered a mutant T1 reovirus incapable of binding the T1 reovirus-specific glycan receptor, GM2. The mutant virus induced substantially less hydrocephalus than wild-type virus, an effect phenocopied by wild-type virus infection of GM2-deficient mice. In comparison to wild-type virus, yields of mutant virus were diminished in cultured ependymal cells, the cell type that lines the brain ventricles. These findings suggest that GM2 engagement targets reovirus to ependymal cells in mice and illuminate the function of glycan engagement in reovirus serotype-dependent disease.ImportanceReceptor utilization strongly influences viral disease, often dictating host range and target cell selection. Different reovirus serotypes bind to different glycans, but a precise function for these molecules in pathogenesis is unknown. We used type 1 (T1) reovirus deficient in binding the GM2 glycan and mice lacking GM2 to pinpoint a role for glycan engagement in hydrocephalus caused by T1 reovirus. This work indicates that engagement of a specific glycan can lead to infection of specific cells in the host and consequent disease at that site. Since reovirus is being developed as a vaccine vector and oncolytic agent, understanding reovirus-glycan interactions may allow manipulation of reovirus glycan-binding properties for therapeutic applications.

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A Single Point Mutation, Asn₁₆?Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

Publications

A Single Point Mutation, Asn<sub>16</sub>→Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

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Dataset Information

A Single Point Mutation, Asn16?Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

Publications

A Single Point Mutation, Asn<sub>16</sub>→Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

A Single Point Mutation, Asn₁₆?Lys, Dictates the Temperature-Sensitivity of the Reovirus tsG453 Mutant.