Cloning and analysis of the CD18 promoter.
CD18, the common beta chain of the leukocyte integrin adhesion proteins, is expressed exclusively by myeloid cells and lymphocytes. During myeloid differentiation, the increase in CD18 cell surface expression is paralleled by increased CD18 messenger RNA levels. Nuclear run-on studies show that CD18 expression is transcriptionally regulated during 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced HL-60 monocytic differentiation. The CD18 transcriptional start site was defined by primer extension and RNAse protection. The gene encoding CD18 was cloned and a fragment that overlaps the transcriptional start site was isolated. DNA sequence analysis of this promoter fragment identified potential AP-1 elements that may mediate the TPA transcriptional response. The CD18 promoter also contains a putative binding site for PU.1, a leukocyte-specific transcription factor. DNA elements resembling those found in other myeloid and integrin promoters were identified. The CD18 promoter fragment was linked to the luciferase reporter gene, electroporated into the U937 monocytic cell line, and its expression increased after exposure to TPA. Thus, CD18 may serve as a model for identifying the cis elements and trans-acting factors that regulate gene expression during myeloid differentiation.
Department of Medicine, Miriam Hospital, Boston, MA.
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D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D018821:CD18 Antigens; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D003001:Cloning, Molecular / Q000379:methods; D015342:DNA Probes; D015698:Genomic Library; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D016028:Receptors, Leukocyte-Adhesion / Q000235:genetics*; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
D015703:Antigens, CD; D018821:CD18 Antigens; D015342:DNA Probes; D009838:Oligodeoxyribonucleotides; D012334:RNA, Neoplasm; D016028:Receptors, Leukocyte-Adhesion; D013755:Tetradecanoylphorbol Acetate