TY - JOUR
T1 - Human β-globin locus control region
T2 - Analysis of the 5′ DNase I hypersensitive site HS 2 in transgenic mice
AU - Caterina, John J.
AU - Ryan, Tom M.
AU - Pawlik, Kevin M.
AU - Palmiter, Richard D.
AU - Brinster, Ralph L.
AU - Behringer, Richard R.
AU - Townes, Tim M.
PY - 1991
Y1 - 1991
N2 - The human β-globin locus control region (LCR) is essential for high-level expression of human ε-, γ-, and β-globin genes. Developmentally stable DNase I hypersensitive sites (designated HS) mark sequences within this region that are important for LCR activity. A 1.9-kilobase (kb) fragment containing the 5′ HS 2 site enhances human β-globin gene expression 100-fold in transgenic mice and also confers position-independent expression. To further define important sequences within this region, deletion mutations of the 1.9-kb fragment were introduced upstream of the human β-globin gene, and the constructs were tested for activity in transgenic mice. Although enhancer activity was gradually lost with deletions of both 5′ and 3′ sequences, a 373-base-pair (bp) fragment retained the ability to confer relative position-independent expression. Three prominent DNase I footprints were observed in this region with extracts from the human erythroleukemia cell line K-562, one of which contained duplicated binding sites for transcription factor AP-1 (activator protein 1). When the 1.9-kb fragment containing an 18-bp deletion of the AP-1 binding sites was tested in transgenic mice, enhancer activity decreased 20-fold but position-independent expression was retained.
AB - The human β-globin locus control region (LCR) is essential for high-level expression of human ε-, γ-, and β-globin genes. Developmentally stable DNase I hypersensitive sites (designated HS) mark sequences within this region that are important for LCR activity. A 1.9-kilobase (kb) fragment containing the 5′ HS 2 site enhances human β-globin gene expression 100-fold in transgenic mice and also confers position-independent expression. To further define important sequences within this region, deletion mutations of the 1.9-kb fragment were introduced upstream of the human β-globin gene, and the constructs were tested for activity in transgenic mice. Although enhancer activity was gradually lost with deletions of both 5′ and 3′ sequences, a 373-base-pair (bp) fragment retained the ability to confer relative position-independent expression. Three prominent DNase I footprints were observed in this region with extracts from the human erythroleukemia cell line K-562, one of which contained duplicated binding sites for transcription factor AP-1 (activator protein 1). When the 1.9-kb fragment containing an 18-bp deletion of the AP-1 binding sites was tested in transgenic mice, enhancer activity decreased 20-fold but position-independent expression was retained.
KW - DNase I cleavage protection patterns
KW - activator protein 1 binding sites
KW - footprinting
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M3 - Article
C2 - 2000371
AN - SCOPUS:0026093060
SN - 0027-8424
VL - 88
SP - 1626
EP - 1630
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 5
ER -