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A gene encoding a novel RFX-associated transactivator is mutated in the majority of MHC class II deficiency patients

Nature Genetics volume 20, pages 273–277 (1998)Cite this article

Abstract

Major histocompatibility class II (MHC-II) molecules are transmembrane proteins that have a central role in development and control of the immune system. They are encoded by a multi-gene family and their expression is tightly regulated. MHC-II deficiency (OMIM 209920) is an autosomal recessive immunodeficiency syndrome resulting from defects in trans-acting factors essential for transcription of MHC-II genes1,2. There are four genetic complementation groups3,4,5 (A, B, C and D), reflecting the existence of four MHC-II regulators2. The factors defective in groups A (CIITA), C (RFX5) and D (RFXAP) have been identified6,7,8. CIITA is a non-DNA-binding co-activator that controls the cell-type specificity and inducibility of MHC-II expression6,9. RFX5 and RFXAP are two subunits of RFX, a multi-protein complex that binds the X box motif of MHC-II promoters10,11. Mutations in the genes encoding RFX5 (RFX5) or RFXAP (RFXAP) abolish binding of RFX (Refs 7,12). Similar to groups C and D, group B is characterized by a defect in RFX binding2,10,11, and although it accounts for the majority of patients, the factor defective in group B has remained unknown. We report here the isolation of RFX by a novel single-step DNA-affinity purification approach and the identification of RFXANK, the gene encoding a third subunit of RFX. RFXANK restores MHC-II expression in cell lines from patients in group B and is mutated in these patients. RFXANK contains a protein-protein interaction region consisting of three ankyrin repeats. Its interaction with RFX5 and RFXAP is essential for binding of the RFX complex to MHC-II promoters.

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Figure 1: Purification of MHC-II promoter-binding proteins and identification of a novel 33-kD subunit of the RFX complex.
Figure 2: Amino acid sequence alignment between RFXANK and homologous proteins containing ankyrin repeats.
Figure 3: RFXANK restored MHC-II expression in cells from patients from complementation group B.
Figure 4: RFXANK mutations in three patients in MHC-II deficiency group B.
Figure 5: A functional RFX complex can be reconstituted with in vitro-translated subunits.
Figure 6: Elucidation of the molecular defects in MHC-II deficiency led to the identification of four essential and specific transactivators of MHC-II expression.

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Acknowledgements

We thank V. Rouge for technical assistance in two-dimensional electrophoretic analysis, R. Mantovani for generous gift of anti-NF-Y antibodies, J. Lee and M. Hadam for providing cells and DNA from the families of patients BLS1 and Ab. We also thank B. Durand for help in the purification of RFX, S. Bontron for providing RNA samples, I. Braun for RNA samples and for help in modelling the RFXANK protein and M. Strubin for helpful discussions and critical reading of the manuscript. This work was supported by grants from the Louis Jeantet Foundation, the Swiss National Science Foundation, and grants from the National Science Foundation (USA) and National Institutes of Health to R.A.

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Authors and Affiliations

  1. Department of Genetics and Microbiology, Louis Jeantet Laboratory of Molecular Genetics, University of Geneva Medical School, 1 rue Michel-Servet, Geneva, 1211, Switzerland

    Krzysztof Masternak, Emmanuèle Barras, Madeleine Zufferey, Bernard Conrad, Bernard Mach & Walter Reith

  2. Department of Molecular Biotechnology, University of Washington, Seattle, 98195-7730, Washington, USA

    Garry Corthals & Ruedi Aebersold

  3. Central Clinical Chemistry Laboratory, Geneva University Hospital, Geneva, 1211, Switzerland

    Jean-Charles Sanchez & Denis F. Hochstrasser

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  1. Krzysztof Masternak
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Correspondence to Bernard Mach or Walter Reith.

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Masternak, K., Barras, E., Zufferey, M. et al. A gene encoding a novel RFX-associated transactivator is mutated in the majority of MHC class II deficiency patients. Nat Genet 20, 273–277 (1998). https://doi.org/10.1038/3081

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