Glycosylation contributes to variability in expression of murine cytomegalovirus m157 and enhances stability of interaction with the NK-cell receptor Ly49H

Natalya V Guseva; Colleen A Fullenkamp; Paul W Naumann; Michael R Shey; Zuhair K Ballas; Jon C D Houtman; Catherine A Forbes; Anthony A Scalzo; Jonathan W Heusel

doi:10.1002/eji.200940134

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Glycosylation contributes to variability in expression of murine cytomegalovirus m157 and enhances stability of interaction with the NK-cell receptor Ly49H

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Glycosylation contributes to variability in expression of murine cytomegalovirus m157 and enhances stability of interaction with the NK-cell receptor Ly49H

Natalya V Guseva, Colleen A Fullenkamp, Paul W Naumann, Michael R Shey, Zuhair K Ballas, Jon C D Houtman, Catherine A Forbes, Anthony A Scalzo and Jonathan W Heusel

European journal of immunology, Vol.40(9), pp.2618-2631

09/2010

DOI: 10.1002/eji.200940134

PMCID: PMC3070389

PMID: 20662096

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Abstract

NK cell-mediated resistance to murine cytomegalovirus (MCMV) is controlled by allelic Ly49 receptors, including activating Ly49H (C57BL/6 strain) and inhibitory Ly49I (129 strain), which specifically recognize MCMV m157, a glycosylphosphatidylinositol-linked protein with homology to MHC class I. Although the Ly49 receptors retain significant homology to classic carbohydrate-binding lectins, the role of glycosylation in ligand binding is unclear. Herein, we show that m157 is expressed in multiple, differentially N-glycosylated isoforms in m157-transduced or MCMV-infected cells. We used site-directed mutagenesis to express single and combinatorial asparagine (N)-to-glutamine (Q) mutations at N178, N187, N213, and N267 in myeloid and fibroblast cell lines. Progressive loss of N-linked glycans led to a significant reduction of total cellular m157 abundance, although all variably glycosylated m157 isoforms were expressed at the cell surface and retained the capacity to activate Ly49H(B6) and Ly49I(129) reporter cells and Ly49H(+) NK cells. However, the complete lack of N-linked glycans on m157 destabilized the m157-Ly49H interaction and prevented physical transfer of m157 to Ly49H-expressing cells. Thus, glycosylation on m157 enhances expression and binding to Ly49H, factors that may impact the interaction between NK cells and MCMV in vivo where receptor-ligand interactions are more limiting.

Protein Binding - genetics

Viral Proteins - immunology

Killer Cells, Natural - pathology

Viral Proteins - metabolism

Lymphocyte Activation - genetics

Myeloid Cells - immunology

Herpesviridae Infections - metabolism

Killer Cells, Natural - immunology

Herpesviridae Infections - genetics

Fibroblasts - metabolism

NK Cell Lectin-Like Receptor Subfamily A - metabolism

Cell Line

Mutagenesis, Site-Directed

Transgenes - genetics

Viral Proteins - genetics

Muromegalovirus - pathogenicity

Glycosylation

Fibroblasts - pathology

Herpesviridae Infections - virology

Mutation - genetics

NK Cell Lectin-Like Receptor Subfamily A - immunology

Muromegalovirus - immunology

Animals

Myeloid Cells - metabolism

Fibroblasts - immunology

Mice

Killer Cells, Natural - metabolism

Myeloid Cells - pathology

Herpesviridae Infections - immunology

Protein Isoforms - genetics

Details

Title: Subtitle: Glycosylation contributes to variability in expression of murine cytomegalovirus m157 and enhances stability of interaction with the NK-cell receptor Ly49H
Creators: Natalya V Guseva - Department of Pathology, Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA 52242, USA
Colleen A Fullenkamp
Paul W Naumann
Michael R Shey
Zuhair K Ballas
Jon C D Houtman
Catherine A Forbes
Anthony A Scalzo
Jonathan W Heusel
Resource Type: Journal article
Publication Details: European journal of immunology, Vol.40(9), pp.2618-2631
DOI: 10.1002/eji.200940134
PMID: 20662096
PMCID: PMC3070389
ISSN: 0014-2980
eISSN: 1521-4141
Grant note: R56 AI063226-01 / NIAID NIH HHS R56 AI063226 / NIAID NIH HHS
Language: English
Date published: 09/2010
Academic Unit: Microbiology and Immunology; Pathology; Immunology; Internal Medicine
Record Identifier: 9984094398002771

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