Copper and the ACE1 regulatory protein reversibly induce yeast metallothionein gene transcription in a mouse extract.

Abstract:

:We describe a cell-free system in which the transcription of the yeast metallothionein gene is inducible by the addition of metal ions plus a specific regulatory protein. Efficient transcription requires the complete yeast ACE1 metalloregulatory protein, including both its DNA-binding and transactivation domains; a mouse nuclear extract providing RNA polymerase and general transcription factors; a template containing the ACE1 binding site; and Cu(I). Because the binding of ACE1 to DNA is dependent on Cu, it is possible to inhibit transcription by the use of Cu-complexing agents such as CN-. We have used this specific inhibition to show that the ACE1 regulatory protein is required for the maintenance as well as the formation of a functional preinitiation complex. The ability to reversibly induce yeast metallothionein gene transcription in vitro provides a powerful system for determining the molecular mechanism of a simple eukaryotic regulatory circuit.

authors

Culotta VC,Hsu T,Hu S,Fürst P,Hamer D

doi

10.1073/pnas.86.21.8377

subject

Has Abstract

pub_date

1989-11-01 00:00:00

pages

8377-81

issue

21

eissn

0027-8424

issn

1091-6490

journal_volume

86

pub_type

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