Article: article from journal or magazin.
Functional domains of the heavy metal-responsive transcription regulator MTF-1
Nucleic Acids Research
Journal Article --- Old month value: Jun 25
Metallothioneins (MTs) constitute a class of low molecular weight, cysteine-rich, metal binding proteins which are regulated at the level of gene transcription in response to heavy metals and other adverse treatments. We have previously cloned a zinc finger factor (MTF-1) that binds specifically to heavy metal-responsive DNA sequence elements in metallothionein promoters and shown that this factor is essential for basal and heavy metal-induced transcription. Here we report that the C-terminal part of MTF-1 downstream of the DNA binding zinc fingers harbours three different transactivation domains, namely an acidic domain, a proline-rich domain and a domain rich in serine and threonine. When fused to the heterologous DNA binding domain of the yeast factor GAL4 these activation domains function constitutively, i.e. transcription of a GAL4-driven reporter gene is not induced by heavy metals. In search of the region(s) responsible for metal induction, external and internal deletion mutations of mouse and human MTF-1 and chimeric variants thereof were tested with a reporter gene driven by a metal-responsive promoter. The N-terminal part of MTF-1 containing the zinc fingers, which are dependent on zinc for efficient DNA binding, can indeed confer a limited (3- to 4-fold) zinc-responsive transcription when fused to the heterologous activation domain of the viral VP16 protein. Another region containing the acidic and proline-rich activation domains also contributes to metal inducibility, but only in the context of intact MTF-1. This indicates that the activity of MTF-1 results from a complex interplay of different functional domains.
Base Sequence Binding Sites DNA/*metabolism DNA-Binding Proteins Fungal Proteins/chemistry/genetics/metabolism Gene Deletion Hela Cells Herpes Simplex Virus Protein Vmw65/chemistry/genetics Humans Hydrogen-Ion Concentration Metallothionein/*genetics Metals/*pharmacology Molecular Sequence Data Mutagenesis Plasmids Proline/analysis Promoter Regions (Genetics) Recombinant Fusion Proteins/chemistry/metabolism *Saccharomyces cerevisiae Proteins Serine/analysis Threonine/analysis Trans-Activation (Genetics) Transcription Factors/*chemistry/genetics/metabolism Zinc Fingers
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