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Isolation and characterization of mouse FM3A cell mutants which are devoid of Newcastle disease virus receptors
T Hara1, S Hattori, M Kawakita
1Department of Pure and Applied Sciences, College of Arts and Sciences, University of Tokyo, Japan.
Abstract:
A method was developed to select host cell mutants which did not permit the replication of Newcastle disease virus (NDV), and 14 isolates of NDV-nonpermissive mutants of mouse FM3A cells were obtained. All these isolates were judged to be deficient in NDV receptors, since their ability to adsorb 3H-labeled NDV virions was markedly decreased. They were tested for genetic complementation in pairs by cell fusion and shown to fall into a single recessive complementation group, which was designated as Had-1. Vesicular stomatitis virus was able to replicate in this mutant to produce infectious progeny, but the glycoprotein of the released virion was abnormal in size, suggesting a defective processing of the asparagine-linked carbohydrate chains in the mutant cell. The Had-1 mutant was resistant to wheat germ agglutinin, but sensitive to a Griffonia simplicifolia lectin, GS-II, which recognizes terminal N-acetylglucosamine residues. The altered sensitivity to these plant lectins compared with that of the parental FM3A cells indicates that sialylated sugar chains on the cell surface are almost absent from the Had-1 cells, thereby rendering the cells NDV receptor deficient.
Insights
Researchers identified mouse cell mutants lacking Newcastle disease virus (NDV) receptors. These Had-1 mutants demonstrate a defect in cell surface sialic acid, preventing NDV entry and replication.
Area of Science:
- Virology
- Cell Biology
- Glycobiology
Background:
- Newcastle disease virus (NDV) requires specific host cell receptors for entry and replication.
- Understanding host-pathogen interactions is crucial for developing antiviral strategies.
Purpose of the Study:
- To develop a method for selecting host cell mutants resistant to NDV.
- To characterize the genetic and molecular basis of NDV non-permissiveness in host cells.
Main Methods:
- Selection of mouse FM3A cell mutants resistant to NDV replication.
- Adsorption assays using radiolabeled NDV virions.
- Genetic complementation analysis via cell fusion.
- Vesicular stomatitis virus replication assays.
- Lectins binding assays (wheat germ agglutinin, Griffonia simplicifolia lectin II).
Main Results:
- 14 NDV-nonpermissive mouse cell mutants were isolated.
- Mutants showed significantly reduced adsorption of NDV virions, indicating a deficiency in NDV receptors.
- Complementation analysis revealed a single recessive complementation group, designated Had-1.
- Had-1 mutants supported vesicular stomatitis virus replication, but produced virions with abnormally sized glycoproteins.
- Mutants exhibited altered lectin binding profiles, suggesting a near absence of cell surface sialylated sugar chains.
Conclusions:
- The Had-1 mutation confers resistance to NDV by impairing cell surface receptor function.
- The mutation affects the processing of asparagine-linked carbohydrate chains, leading to altered cell surface glycans.
- This study identifies a specific host cell defect crucial for NDV entry.