In vitro recognition of specific DNA targets by AlcR, a zinc binuclear cluster activator different from the other proteins of this class.

The Journal of Biological Chemistry
F LenouvelB Felenbok

Abstract

AlcR is the transactivator mediating transcriptional induction of the alc gene cluster in Aspergillus nidulans. The AlcR DNA-binding domain consists of a zinc binuclear cluster different from the other members of the Zn2Cys6 family by several features. In particular, it is able to bind to symmetric and asymmetric sites with the same affinity, with both sites being functional in A. nidulans. Here, we show that unlike the other proteins of the Zn2Cys6 binuclear cluster family, AlcR binds most probably as a monomer to its cognate targets. Two molecules of the AlcR protein can simultaneously bind in a noncooperative manner to inverted repeats. The consensus core has been determined precisely (5'-CCGCN-3'), and the AlcR-binding site in the aldA promoter has been localized. The sequence downstream of the zinc cluster is necessary for high affinity binding. Furthermore, our data show that the use of the carrier protein glutathione S-transferase in AlcR binding experiments introduces an important bias in the recognition of DNA sites due to its tertiary dimeric structure.

References

Apr 2, 1992·Nature·R MarmorsteinS C Harrison
Dec 28, 1969·Journal of Molecular Biology·M E Goldberg
Jan 1, 1980·Methods in Enzymology·A M Maxam, W Gilbert
Aug 15, 1995·Proceedings of the National Academy of Sciences of the United States of America·H E XuS A Johnston
Jun 30, 1994·Nature·R H JacobsonB W Matthews
Sep 1, 1993·Molecular and Cellular Biology·R M Brazas, D J Stillman
Jun 21, 1996·Journal of Molecular Biology·J TimmermanB Guiard

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Citations

May 23, 2001·Archives of Biochemistry and Biophysics·J M RheeE E Turner
Sep 12, 2002·Fungal Genetics and Biology : FG & B·I NikolaevB Felenbok
Aug 16, 2003·European Journal of Biochemistry·Michel FlipphiBéatrice Felenbok
Mar 10, 2006·Eukaryotic Cell·Roman RauscherRobert L Mach
Jul 16, 2003·Molecular and Cellular Biology·Jeong-Ho KimMark Johnston
Jul 11, 1998·The Journal of Biological Chemistry·Y MamaneB Turcotte
Dec 16, 1998·The Journal of Biological Chemistry·J M RheeE E Turner
Jul 4, 1998·The Journal of Biological Chemistry·J Noël, B Turcotte
Oct 6, 1998·Applied and Environmental Microbiology·N N van PeijL H de Graaff
Mar 30, 2001·Fungal Genetics and Biology : FG & B·I G JonesH M Sealy-Lewis

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