PMID: 2497463May 1, 1989Paper

Structure and function of the Zn(II) binding site within the DNA-binding domain of the GAL4 transcription factor

Proceedings of the National Academy of Sciences of the United States of America
T Pan, J E Coleman

Abstract

The transcription factor GAL4 from Saccharomyces cerevisiae contains a "zinc-finger"-like motif, Cys-Xaa2-Cys-Xaa6-Cys-Xaa6-Cys-Xaa2-Cys-Xaa6+ ++-Cys, within its DNA-binding domain. A GAL4 fragment consisting of residues 1-147 plus two additional residues from the cloning vector [denoted GAL4(149*)] has been cloned and overexpressed in Escherichia coli. This fragment includes the entire DNA-binding domain (residues 1-74). The homogeneous GAL4-(149*) protein contains 1-1.5 moles of Zn(II) per mole of protein. The GAL4(149*) protein binds tightly to the specific 17-base-pair palindromic DNA sequence found at GAL4 binding sites as shown by gel-retention assays using a 32P-labeled 23-mer containing this sequence. Removal of the intrinsic Zn(II) by EDTA at low pH abolishes binding to the 23-mer. The GAL4(149*) apoprotein can be reconstituted with Zn(II), Cd(II), or Co(II) with restoration of specific DNA binding. Titration of GAL4(149*) apoprotein with 113Cd(II) shows two 113Cd(II) binding sites on the molecule, one with delta of 707 ppm, suggesting coordination to four sulfur atoms, and one with delta of 669 ppm, suggesting coordination to three or four sulfur atoms. Because GAL4(149*) protein contains only six cysteine residues wi...Continue Reading

References

Mar 21, 1989·Biochemistry·D P GiedrocJ E Coleman
Aug 25, 1987·Biochemistry·D P GiedrocJ E Coleman
May 5, 1986·Journal of Molecular Biology·F W Studier, B A Moffatt
Apr 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·M Johnston, J Dover

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Citations

May 1, 1992·Protein Science : a Publication of the Protein Society·L A Basile, J E Coleman
Apr 1, 1994·Protein Science : a Publication of the Protein Society·K K Rodgers, J E Coleman
Nov 1, 1994·Journal of Biomolecular NMR·K H Gardner, J E Coleman
Mar 1, 1992·The Journal of Steroid Biochemistry and Molecular Biology·K Dahlman-WrightJ A Gustafsson
Mar 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Pan, J E Coleman
Feb 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·B L ValleeD S Auld
Nov 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·M CzuprynB L Vallee
Mar 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·W T Miller, P Schimmel
Mar 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·R LoveringP S Freemont
May 2, 2007·Bioinformatics·Arijit ChakravartyRobert H Gross
May 13, 1999·Proceedings of the National Academy of Sciences of the United States of America·N R Yaseen, G Blobel
Feb 1, 1995·Medical Hypotheses·R T Radulescu
Mar 15, 1991·Biochemical and Biophysical Research Communications·A HansenT Kodadek
Aug 16, 2016·Bioinorganic Chemistry and Applications·Dongxin Zhao, Zhongxian Huang
Oct 26, 2016·Epigenetics & Chromatin·Laura BruecknerBas van Steensel
Mar 27, 1990·Biochemistry·T Pan, J E Coleman
Feb 9, 1993·Biochemistry·M CzuprynB L Vallee
Oct 1, 1995·Nature Structural Biology·K H GardnerJ E Coleman
Sep 1, 1995·Journal of Biomolecular NMR·B I SchweitzerG Tucker-Kellogg
Jun 9, 2005·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Sang Wook OhEui Yul Choi
May 1, 1991·Molecular and Cellular Biology·J E Marczak, M C Brandriss

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