Nucleic Acids Research, 1991, Vol. 19, No. 6 1243-1249
© 1991
MOLECULAR BIOLOGY |
Regulatory factor-X binding to mutant HLA-DRA promoter sequences
Department of Microbiology and Immunology, Emory University School of Medicine Atlanta, GA 30322, USA 1Department of Microbiology, University of Geneva Geneva, Switzerland
* To whom correspondence should be addressed
Received December 19, 1990. Accepted February 6, 1991.
The class II genes of the major histocompatibility complex (MHC) encode a family of cell surface glycoprotelns that present processed antigen to the T cell receptor. Class II genes are regulated coordinately, responding to both immunologic and developmental signals. Conserved sequence elements 5' to class II genes have been shown to be important in transcriptional control. One of these sequences, the X box, is a specific target for the binding of the factor RF-X. In the hereditary HLA class II deficiency, a form of primary immunodeficiency, a regulatory defect In expression of class II genes is associated with a defect in the binding of RF-X. To determine the basepairs that are important for this binding interaction, a series of single basepalr substitutions spanning the X box motif of the DRA gene was constructed and tested for binding of RF-X by gel electrophoresis mobility shift assays (EMSAS). Several, but not all, of the mutants severely affected binding of RF-X. In addition, the binding of both the natural and the recombinant form of RF-X was affected with the same specificity. A comparison of X box basepair positions important for RF-X binding to DRA with sequences conserved between X boxes of other class II alpha chain genes suggests that high affinity RF-X binding is important for a high level of expression and may explain differences in the levels of class II expression of different class II alpha chains.
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