Nucleotide sequence analysis of H-2Df and the spontaneous in vivo H-2Dfm2 mutation

The nucleotide sequence of the standard H-2Df allele and the spontaneous in vivo H-2Dfm2 mutation are reported here. Locus-specific sequences in the 5' and 3' untranslated regions of the mouse MHC class I H-2D-region genes were used to design primers for the specific amplification and cloning of H-2D-region cDNA from standard B10.M/Sn H-2f and mutant B10.M-H-2fm2/Mob mice. A partial Df genomic clone and direct Df and Dfm2 mRNA sequence analysis confirmed the authenticity of the cDNA clones. Interestingly, H-2Df contains a proline in the alpha-helix of the alpha 1 domain at amino acid position 62; no other known class I molecule has a proline at this position. The H-2Dfm2 mutation, however, replaces this unique proline in Df with the H-2 and HLA consensus arginine at position 62. Although a point mutation cannot be ruled out, the single nucleotide change in the H-2Dfm2 mutation is flanked by a stretch of 47 nucleotide bases with an identical counterpart in H-2Kf, a finding consistent with a recombinatorial event between H-2Kf and H-2Df.