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Nucleic Acids Research, 1987, Vol. 15, No. 20 8399-8415
© 1987

Articles

Tissue-specific expression of the human {alpha}1-antitrypsin gene is controlled by multiple cis-regulatory elements

Rong-Fong Shen1,2, Yi Li2, Richard N. Sifers2, Heng Wang2, Christopher Hardick2, Sophia Y. Tsai2 and Savio L.C. Woo1,2,3

1Howard Hughes Medical Institute Houston, TX 77030, USA 2Department of Cell Biology Houston, TX 77030, USA 3Institute for Molecular Genetics, Baylor College of Medicine Houston, TX 77030, USA

Received July 2, 1987. Revised September 16, 1987. Accepted September 16, 1987.

Human {alpha}1-antitrypsin (AAT) is expressed in the liver, and a 318 bp fragment immediately flanking the CAP site of the gene was found to be sufficient to drive the expression of a reporter gene (CAT) specifically in hepatoma cells. The enhancing activity however, was orientation-dependent. The DNA fragment was separated into a distal region and a proximal region. A "core enhancer" sequence GTGGTTTC is present within the distal region and is capable of activity enhancement in both orientations when complemented by the proximal region in the sense orientation. The results strongly suggest that there are multiple cis-acting elements in the human AAT gene that confer cell specificity for its expression. Nuclear proteins prepared from the hepatoma cells bound specifically to the proximal region in a band-shifting assay that was resistant to competition by the globin promoter DNA. Foot-printing analysis showed a protected domain within the proximal region that contains a nearly perfect palindromic sequence TGGTTAATATTCACCA, which may be important in the regulation of AAT expression in the liver.


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