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Harlow P , Watkins E , Thornton RD , Nemer M
Structure of an ectodermally expressed sea urchin metallothionein gene and characterization of its metal-responsive region
Molecular and Cellular Biology. 1989 ;9(12) :5445-5455
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Abstract
The metallothionein-A gene in the metallothionein gene family of the sea urchin Strongylocentrotus purpuratus (SpMTA gene) was sequenced and found to contain three coding exons plus a 3' entirely noncoding exon. Putative ? and ? MT domains were encoded, by its exons 2 and 3, respectively, in reverse of the order in vertebrate metallothionein genes. The SpMTA promoter was characterized through the expression of recombinant constructs containing various portions of the proximal 678-base-pair (bp) 5'-flanking region of the SpMTA gene. Zygotes injected with constructs were cultured to the blastula stage in the presence of a heavy-metal chelator and then incubated in the presence or absence of cadmium. The longest constructs were expressed only when heavy-metal ion was present. Two putative metal-responsive elements (MREs a and b) within 240 bp of the transcription start site resembled mammalian MREs in their critical 8-bp cores (TGCRCNCS) and in their locations relative to each other and to the TATA box. Elimination of activity by site-specific mutations in MREs a and b, separately or in both, identified them as metal regulatory elements. Thus, MRE recognition in this invertebrate resembles that in vertebrates. Upstream sites with single-mismatched MREs neither acted as MREs nor amplified the activity of MREs a and b. The SpMTA, Spec1, and CyIIIa actin genes, which have the same ectodermal specificity, have common DNA elements at relatively similar locations in their promoter regions; however, these elements are insufficient in themselves to promote gene expression.
Notes
02707306 (ISSN) Cited By: 12; Export Date: 30 May 2006; Source: Scopus CODEN: MCEBD Language of Original Document: English Correspondence Address: Nemer, M.; Institute for Cancer Research; Fox Chase Cancer Center; 7701 Burholme Avenue Philadelphia, PA 19111, United States Molecular Sequence Numbers: GENBANK: M30606; Chemicals/CAS: Metallothionein, 9038-94-2; Metals