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Identification and sequence analysis of the promoter for the leukocyte
integrin beta-subunit (CD18): a retinoic acid-inducible gene
ED Agura, M Howard and SJ Collins
Molecular Medicine Program, Fred Hutchinson Cancer Research Center,
Seattle, WA 98104.
Leukocyte adhesion receptors (LFA-1; Mac-1; p150,95) are a family of
heterodimeric cell-surface adhesion molecules expressed exclusively in
granulocytes, lymphocytes, and macrophages. Expression of these proteins is
under complex regulatory control, but to date promoters for these genes
have not been identified. The CD18 gene codes for the common beta-subunit
of the leukocyte adhesion receptors. Transcription of CD18 is highly
tissue-specific, hormonally inducible (by retinoic acid [RA]), and
coordinately regulated with leukocyte integrin alpha- chains. To identify
the CD18 promoter, we screened a human genomic phage library with a human
CD18 cDNA probe and obtained a clone that contains an exon coding for the
5' untranslated region (UTR). Using rapid amplification of cDNA ends
(RACE), RNAse protection, S1 nuclease, and primer extension assays, we
demonstrated the existence of multiple transcription start sites clustered
in a 45-nt region. We investigated the transcription-promoting activity of
the genomic sequences 5' to the CD18 gene by performing transient
expression assays with a growth hormone reporter gene in various
hematopoietic cell lines. The CD18 promoter was active in Jurkat cells, a
lineage that normally expresses CD18 but was considerably less active in
K562, an early erythroid line that does not normally express CD18. The
genomic sequences upstream of the start site cluster lack CAAT and TATA
boxes, but have two Sp1 binding sites and 10 T(G/C)AC(C/A) boxes, which may
represent binding sites for RA receptors (RAR). These features distinguish
the CD18 promoter from the promoters of other tissue-specific,
hormone-inducible genes, and may be representative of leukocyte integrin
promoters in general.
Volume 79,
Issue 3,
pp. 602-609,
02/01/1992
Copyright © 1992 by The American Society of Hematology
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