Aberrant splicing of intron 1 creates a novel null HLA-B*1501 allele
| Autor: | M D, Curran, F, Williams, A M, Little, B K, Rima, J A, Madrigal, D, Middleton |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Base Sequence
Histocompatibility Testing Blotting Western Molecular Sequence Data HLA-B15 Antigen Polymerase Chain Reaction Introns Tissue Donors Alternative Splicing Open Reading Frames Bone Marrow HLA-B Antigens Humans Amino Acid Sequence RNA Messenger Registries Isoelectric Focusing Alleles Sequence Deletion |
| Zdroj: | Tissue antigens. 53(3) |
| ISSN: | 0001-2815 |
| Popis: | A comparison of serological and DNA HLA class I typing data identified a serological "blank" HLA-B15 antigen in a volunteer donor on the bone marrow registry. Isoelectric focusing and Western blot analysis of a cell line established from this individual confirmed that the HLA-B15 antigen is not expressed at the cell surface. Nucleotide sequence analysis of the HLA-B*15 null allele revealed a 10-bp deletion near the 3' end of intron 1, when compared to the normal HLA-B*1501 sequence. All of the HLA-B*15 specific cDNA clones examined retained the intron 1 sequence. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis demonstrated that the HLA-B*15 mRNA molecule contained the intron 1 sequence, indicating an inability to efficiently splice out intron 1 from the mRNA transcript. The retention of the mutated intron 1 sequence in the mRNA causes a frameshift and premature termination of translation at the start of exon 2, explaining the HLA-B*1501 null phenotype. Our data predicts that the HLA-B*1501 null allele would express a small truncated protein containing the signal sequence fused to an ORF within intron 1 and terminating (out of frame) just within exon 2. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|