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Nucleic Acids Research, 1992, Vol. 20, No. 11 2755-2760
© 1992

MOLECULAR BIOLOGY

Synergistic induction of promoters containing metal- and glucocorticoid-responsive elements

Jorge Filmus*, Jacek Remani and Michel H. Klein

The Connaught Centre for Biotechnology Research 1755 Steeles Avenue West, Willowdale, Ontario M2R 3T4, Canada

*To whom correspondence should be addressed at Sunnybrook Health Science Centre, Division of Cancer Research, Reichmann Research Building, S-218, 2075 Bayview Avenue, Toronto, Ontario M4N 3M5, Canada

Received February 26, 1992. Revised April 24, 1992. Accepted April 24, 1992.

We have shown that it is possible to synergistically activate gene transcription when several glucocorticoid responsive elements (GREs) and metal responsive elements (MREs) that coexist within the same promoter are induced simultaneously. To demonstrate this, additional GREs were introduced Into a human metallothionein hA (hMT-IIA) promoter In which some constitutive elements had been deleted. The transcriptional strength and inducibility of the modified hMT-IIA promoters were studied in transient expression experiments using the CAT gene as a reporter. As result of synergistic activation of transcription by CdCl2 and dexamethasone, the induced expression levels the modified promoters were significantly higher than those obtained with wild-type hMT-IIA. Since the increase in inducible expression was not accompanied by a concominant increase in basal levels, the inducibility of the modified MT promoters was up to 6-fold higher. The degree of transcriptional synergism was shown to depend on the position and the number of GREs introduced. Thus, the engineering of synthetic promoters that include both GREs and MREs should offer the opportunity to develop a new series of improved inducible mammalian expression vectors.


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