Upstream DNA Sequences Required for Tissue-Specific Expression of the HLA-DRα Gene

We have used in vitro deletion mutagenesis in combination with DNA transfection to search for cis-acting regulatory elements involved in the tissue-specific expression of a human class II major histocompatibility complex gene. A 140-base-pair 5′ flanking fragment that contains the class II box conse...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-06, Vol.84 (12), p.4254-4258
Main Authors: Sherman, Paula A., Basta, Patricia V., Jenny P.-Y. Ting
Format: Article
Language:English
Subjects:
B lymphocytes
B-Lymphocytes - immunology
Base Sequence
Biological and medical sciences
Cell Line
Cell lines
DNA
Fundamental and applied biological sciences. Psychology
Gels
Gene expression
Genes
Genes, Regulator
HeLa cells
HLA-D Antigens - genetics
HLA-DR Antigens - genetics
Humans
Molecular and cellular biology
Molecular genetics
Mutation
Oligonucleotide probes
Oligonucleotides
Plasmids
Promoter Regions, Genetic
T lymphocytes
T-Lymphocytes - immunology
Transcription, Genetic
Transfection
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Summary:We have used in vitro deletion mutagenesis in combination with DNA transfection to search for cis-acting regulatory elements involved in the tissue-specific expression of a human class II major histocompatibility complex gene. A 140-base-pair 5′ flanking fragment that contains the class II box consensus sequences and an octamer sequence (AT-TTGCAT) confers tissue specificity on the promoter of the HLA-DRα gene. Recombinant DNA plasmids containing this DRα gene segment fused to the coding sequence of the bacterial chloramphenicol acetyltransferase gene are expressed at higher levels in human B-cell lines than in human T-cell lines. We have demonstrated that the most 5′ of the class II boxes is essential for tissue-specific DRα promoter function. In addition, using an electrophoretic mobility shift assay to identify DNA binding proteins, we have detected binding of nuclear proteins to DNA probes containing the class II boxes and the octamer sequence. A protein that binds to the octamer is present at higher levels in nuclear extracts of B-cell lines than in other cell lines examined. This protein may be important for the tissue-specific expression of the HLA-DRα gene.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.12.4254