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Molecular and Cellular Biology, February 1999, p. 1558-1568, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Localization of Distant Urogenital System-, Central Nervous System-, and Endocardium-Specific Transcriptional Regulatory Elements in the GATA-3 Locus

Ganesh Lakshmanan,1 Ken H. Lieuw,1 Kim-Chew Lim,1 Yi Gu,1 Frank Grosveld,2 James Douglas Engel,1,* and Alar Karis2,3

Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 602081; Department of Cell Biology and Genetics, Erasmus University School of Medicine, Rotterdam 3000, Holland2; and Institute of Molecular and Cell Biology, University of Tartu, Tartu EE2400, Estonia3

Received 2 September 1998/Returned for modification 8 October 1998/Accepted 26 October 1998

We found previously that neither a 6-kbp promoter fragment nor even a 120-kbp yeast artificial chromosome (YAC) containing the whole GATA-3 gene was sufficient to recapitulate its full transcription pattern during embryonic development in transgenic mice. In an attempt to further identify tissue-specific regulatory elements modulating the dynamic embryonic pattern of the GATA-3 gene, we have examined the expression of two much larger (540- and 625-kbp) GATA-3 YACs in transgenic animals. A lacZ reporter gene was first inserted into both large GATA-3 YACs. The transgenic YAC patterns were then compared to those of embryos bearing the identical lacZ insertion in the chromosomal GATA-3 locus (creating GATA-3/lacZ "knock-ins"). We found that most of the YAC expression sites and tissues are directly reflective of the endogenous pattern, and detailed examination of the integrated YAC transgenes allowed the general localization of a number of very distant transcriptional regulatory elements (putative central nervous system-, endocardium-, and urogenital system-specific enhancers). Remarkably, even the 625-kbp GATA-3 YAC, containing approximately 450 kbp and 150 kbp of 5' and 3' flanking sequences, respectively, does not contain the full transcriptional regulatory potential of the endogenous locus and is clearly missing regulatory elements that confer tissue-specific expression to GATA-3 in a subset of neural crest-derived cell lineages.

* Corresponding author. Mailing address: Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, 2153 N. Campus Dr., Evanston, IL 60208-3500. Phone: (847) 491-5139. Fax: (847) 467-2152. E-mail: d-engel{at}nwu.edu.

Molecular and Cellular Biology, February 1999, p. 1558-1568, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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