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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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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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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DOI: https://doi.org/10.1038/3081
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