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Volume 272, Number 4, Issue of January 24, 1997 pp. 2334-2341
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Diverse Genetic Regulatory Motifs Required for Murine Adenosine Deaminase Gene Expression in the Placenta

(Received for publication, August 7, 1996)

Daqing Shi Dagger , John H. Winston Dagger , Michael R. Blackburn Dagger , Surjit K. Datta Dagger , Gerri Hanten Dagger and Rodney E. Kellems Dagger **

From the Dagger  Verna and Marrs McLean Department of Biochemistry and ** Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030

Murine adenosine deaminase (ADA) is a ubiquitous purine catabolic enzyme whose expression is subject to developmental and tissue-specific regulation. ADA is enriched in trophoblast cells of the chorioallantoic placenta and is essential for embryonic and fetal development. To begin to understand the genetic pathway controlling Ada gene expression in the placenta, we have identified and characterized a 770-base pair fragment located 5.4 kilobase pairs upstream of the Ada transcription initiation site, which directs reporter gene expression to the placenta of transgenic mice. The expression pattern of the reporter gene reflected that of the endogenous Ada gene in the placenta. Sequence analysis revealed potential binding sites for bHLH and GATA transcription factors. DNase I footprinting defined three protein binding regions, one of which was placenta-specific. Mutations in the potential protein binding sites and footprinting regions resulted in loss of placental expression in transgenic mice. These findings indicate that multiple protein binding motifs are necessary for Ada expression in the placenta.


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