Expression of mutant H-2(d) proteins encoded by class I genes which alternatively process the 5' end of their transcripts

M. L. Hedley, K. Ozato, J. Maryanski, P. W. Tucker, J. Forman

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Mutation of the 3' splice sites bordering exon 2 of the H-2D(d) and H-2K(d) genes generated alternatively spliced transcripts when the constructs were transfected into L cells (J. Immunol. 143:1018). The H-2D(d) transcripts contained an additional 84 nucleotides derived from the first intervening sequence, whereas 60 extra bases were included in the H-2K(d) mRNA. Proteins derived from these transcripts were recognized by mAb. Moreover, both Ag served as recognition elements for CTL, and the mutant H-2K(d) molecule functioned as a restricting element for an Ag peptide. As a result of alternative splicing, the mutant proteins should have additional residues at their NH2 termini to increase their lengths by 28 (D(d)) or 20 (K(d)) amino acids. Immunoprecipitation and analysis on SDS-PAGE demonstrated that the mutant H-2K(d) molecule was indeed larger than the normal H-2K(d) protein, but the mutant and wild-type H-2D(d) Ag were the same size. In addition, treatment of H-2D(d) mutant and normal Ag with N-glycanase produced molecules of equal size, demonstrating that the mutant protein was completely glycosylated. Limited amino acid sequencing of this Ag indicated that it was normal H-2D(d). Therefore, before its transfer to the cell surface, post-translational modifications remove the additional NH2-terminal residues of the mutant D(d) but not K(d) protein.

Original languageEnglish (US)
Pages (from-to)1026-1031
Number of pages6
JournalJournal of Immunology
Volume143
Issue number3
StatePublished - Jan 1 1989

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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