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Isolation of Fidelity Variants of RNA Viruses and Characterization of Virus Mutation Frequency
Published on: June 16, 2011
Analysis of the MCMV resistome by ENU mutagenesis
Karine Crozat1, Philippe Georgel, Sophie Rutschmann
1Department of Immunology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, California 92037, USA. bruce@scripps.edu
Abstract:
The mouse cytomegalovirus (MCMV) resistome is the set of host genes with nonredundant functions in resistance to MCMV infection. By screening 3,500 G(3) germline mutant mice ( approximately 1,750 gamete equivalents), we have identified eight transmissible mutations that create MCMV susceptibility in C57BL/6 mice. Among these, a mutation called Domino was noted to cause macrophage susceptibility to vesicular stomatitis virus (VSV) in vitro. This accessory phenotype was not corrected by type I interferon (IFN), which suggested a defect of the type I IFN pathway. Domino corresponds to a point mutation that alters the DNA binding domain of STAT1, leading to a defect of STAT1 activation. Identification of the Domino mutation demonstrates that an in vivo MCMV susceptibility screen is feasible and illustrates how it can provide insight into the resistome. Moreover, some mutations are far more deleterious than Domino in MCMV-infected mice, consistent with the interpretation that certain protein(s) unrelated to IFN production or signaling are more important than IFNs with regard to their net antiviral effects.
Insights
Researchers identified eight mutations causing mouse cytomegalovirus (MCMV) susceptibility by screening mutant mice. One mutation, Domino, revealed a defect in STAT1 activation, impacting the type I interferon pathway and host antiviral defense.
Area of Science:
- Virology
- Immunology
- Genetics
Background:
- The mouse cytomegalovirus (MCMV) resistome comprises host genes crucial for resisting MCMV infection.
- Understanding host-pathogen interactions is vital for developing antiviral strategies.
Purpose of the Study:
- To identify host genes conferring resistance to MCMV infection through a large-scale genetic screen.
- To investigate the functional role of identified mutations in host antiviral defense mechanisms.
Main Methods:
- Screening of 3,500 G(3) germline mutant mice to identify MCMV-susceptible phenotypes.
- In vitro assays using macrophages to assess susceptibility to vesicular stomatitis virus (VSV).
- Genetic and molecular analysis to characterize the Domino mutation and its effect on STAT1 signaling.
Main Results:
- Eight transmissible mutations conferring MCMV susceptibility were identified in C57BL/6 mice.
- The Domino mutation caused macrophage susceptibility to VSV, independent of type I interferon (IFN) response.
- Domino was mapped to a point mutation in STAT1, impairing its DNA binding and activation.
Conclusions:
- In vivo MCMV susceptibility screens are effective for discovering host resistance factors.
- STAT1 activation is critical for antiviral defense against MCMV and VSV.
- Host factors beyond type I IFN signaling play significant roles in controlling MCMV infection.
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