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Nucleic Acids Research, 1994, Vol. 22, No. 7 1226-1233
© 1994

MOLECULAR BIOLOGY

Structure and regulation of the chicken erythroid {delta}-aminolevulinate synthase gene

Kim-Chew Lim, Hajime Ishihara1, Robert D. Riddle2, Zhuoying Yang, Nancy Andrews3, Masayuki Yamamoto1 and James Douglas Engel*

Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University Evanston, IL 60208, USA 1Department of Biochemistry, Tohoku University School of Medicine 2-1 Seiryomachi, Sendai 980, Japan 2Department of Genetics USA 3Division of Hematology/Oncology, Harvard Medical School Boston, MA 02115, USA

* To whom correspondence should be addressed

Received December 14, 1993. Revised March 2, 1994. Accepted March 2, 1994.

Erythroid cells regulate heme biosynthesis in a manner that is distinct from all other cell types. While heme negatively regulates the synthesis of the housekeeping {delta}-amlnolevullnate synthase (ALAS-N) in all non-erythrold cells, the expression of an erythroid-speclfic isozyme (ALAS-E) is developmentally regulated in red blood cells. As a first step towards understanding the molecular basis for the transcrlptional regulation of ALAS-E during erythropolesis, we cloned and characterized the chicken ALAS-E locus. This gene spans 18 kbp and is composed of eleven exons. The intron/exon structure of erythroid ALAS was found to be conserved among several vertebrate species. Direct RNA sequencing identified a 5' untranslated region that is derived from two contiguous exons and is predicted to form a very stable stem-loop structure that bears resemblance to the ferrltln iron-responsive element. Tissue-specific expression of the ALAS-E gene was analyzed by transient transfectlon assays in hematopoietlc cells of both erythroid and non-erythroid origins. These experiments identified distal (–784 to –505 bp) and proximal (–155 to +21 bp) promoter elements which are required for high level, erythroid-speclfic transcription.


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