title: A novel mutation in IFN-gamma receptor 2 with dominant negative activity: biological consequences of homozygous and heterozygous states.
abstract: We identified two siblings homozygous for a single base pair deletion in the IFN-gammaR2 transmembrane domain (791delG) who presented with multifocal Mycobacterium abscessus osteomyelitis (patient 1) and disseminated CMV and Mycobacterium avium complex infection (patient 2), respectively. Although the patients showed no IFN-gammaR activity, their healthy heterozygous parents showed only partial IFN-gammaR activity. An HLA-identical bone marrow transplant from the mother led patient 1 to complete hemopoietic reconstitution, but only partial IFN-gammaR function. We cloned and expressed fluorescent fusion proteins of the wild-type IFN-gammaR2, an IFN-gammaR2 mutant previously described to produce a complete autosomal recessive deficiency (278del2), and of 791delG to determine whether the intermediate phenotype in the 791delG heterozygous state was caused by haploinsufficiency or a dominant negative effect. When cotransfected together with the wild-type vector into IFN-gammaR2-deficient fibroblasts, the fusion protein with 791delG inhibited IFN-gammaR function by 48.7 +/- 5%, whereas fusion proteins with 278del2 had no inhibitory effect. Confocal microscopy of 791delG fusion proteins showed aberrant diffuse intracellular accumulation without plasma membrane localization. The fusion protein created by 791delG did not complete Golgi processing, and was neither expressed on the plasma membrane, nor shed extracellularly. The mutant construct 791delG exerts dominant negative effects on IFN-gamma signaling without cell surface display, suggesting that it is acting on pathways other than those involved in cell surface recognition of ligand.
authors.affiliation: Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Sergio D
authors.initials: SD
authors.lastname: Rosenzweig
authors.forename: Susan E
authors.initials: SE
authors.lastname: Dorman
authors.forename: Gulbu
authors.initials: G
authors.lastname: Uzel
authors.forename: Stephen
authors.initials: S
authors.lastname: Shaw
authors.forename: Amy
authors.initials: A
authors.lastname: Scurlock
authors.forename: Margaret R
authors.initials: MR
authors.lastname: Brown
authors.forename: Rebecca H
authors.initials: RH
authors.lastname: Buckley
authors.forename: Steven M
authors.initials: SM
authors.lastname: Holland
mesh_terms: D002460:Cell Line; D002461:Cell Line, Transformed; D005260:Female; D005799:Genes, Dominant / Q000276:immunology; D005808:Genes, Recessive / Q000276:immunology; D006580:Genetic Carrier Screening / Q000379:methods; D006720:Homozygote; D006801:Humans; D015151:Immunoblotting; D007223:Infant; D007371:Interferon-gamma / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology; D008297:Male; D015269:Mycobacterium avium Complex / Q000276:immunology; D015270:Mycobacterium avium-intracellulare Infection / Q000235:genetics / Q000276:immunology; D010019:Osteomyelitis / Q000235:genetics / Q000276:immunology / Q000382:microbiology; D017471:Receptors, Interferon / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D017384:Sequence Deletion / Q000276:immunology; D014162:Transfection; D000096743:Interferon gamma Receptor
chemical_list: D007371:Interferon-gamma; D017471:Receptors, Interferon; D011993:Recombinant Fusion Proteins; D000096743:Interferon gamma Receptor; C086185:IFNGR2 protein, human