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J. Biol. Chem., Vol. 265, Issue 14, 8285-8296, May, 1990
DS Boam, AR Clark and K Docherty
Tissue-specific expression of the human insulin gene is regulated by
cis-acting DNA elements 5' to the transcription start site. Deletion of the
5' region of the human insulin gene between nucleotides -279 and - 258
caused a 25-fold rise in transcriptional activity whereas further deletion
to nucleotide -229 reduced transcription activity 25-fold. In vitro
analysis of protein binding in the 5' regulatory region revealed: (i) the
major positive regulatory region (-258 to -229) contains a protein-binding
site (GC-II) with 75% sequence identity to a motif in the rat insulin I
gene, shown to be a powerful transcriptional activator. GC-II motif-binding
factors are not restricted to insulin- producing cell lines. (ii) An islet
cell-specific factor binds between nucleotides -217 to -210 (CT-II motif).
(iii) A region between nucleotides -153 and -127, containing two identical
motifs, GG-I and GG- II was also revealed. GG-I-binding factors are
ubiquitous, whereas binding to the GG-II motif is beta cell-specific. (iv)
A ubiquitous factor binds to a motif between nucleotides -179 and -183,
identical to a half-site for the cyclic nucleotide regulatory element. (v)
The negative regulatory element between -279 and -258 contains overlapping
binding sites for at least 3 protein factors, with differing cell- specific
distributions and can independently down-regulate thymidine kinase promoter
activity in a beta cell line.
Positive and negative regulation of the human insulin gene by multiple trans-acting factors
Department of Medicine, University of Birmingham, Queen Elizabeth Hospital, Edgbaston, United Kingdom.
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