The C6 zinc finger and adjacent amino acids determine DNA-binding specificity and affinity in the yeast activator proteins LAC9 and PPR1.

Molecular and Cellular Biology
M M Witte, R C Dickson

Abstract

LAC9 is a DNA-binding protein that regulates transcription of the lactose-galactose regulon in Kluyveromyces lactis. The DNA-binding domain is composed of a zinc finger and nearby amino acids (M. M. Witte and R. C. Dickson, Mol. Cell. Biol. 8:3726-3733, 1988). The single zinc finger appears to be structurally related to the zinc finger of many other fungal transcription activator proteins that contain positively charged residues and six conserved cysteines with the general form Cys-Xaa2-Cys-Xaa6-Cys-Xaa6-9-Cys-Xaa2-Cys-Xaa 6-Cys, where Xaan indicates a stretch of the indicated number of any amino acids (R. M. Evans and S. M. Hollenberg, Cell 52:1-3, 1988). The function(s) of the zinc finger and other amino acids in DNA-binding remains unclear. To determine which portion of the LAC9 DNA-binding domain mediates sequence recognition, we replaced the C6 zinc finger, amino acids adjacent to the carboxyl side of the zinc finger, or both with the analogous region from the Saccharomyces cerevisiae PPR1 or LEU3 protein. A chimeric LAC9 protein, LAC9(PPR1 34-61), carrying only the PPR1 zinc finger, retained the DNA-binding specificity of LAC9. However, LAC9(PPR1 34-75), carrying the PPR1 zinc finger and 14 amino acids on the carboxyl sid...Continue Reading

References

Mar 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·T Pan, J E Coleman
Jan 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·E Giniger, M Ptashne
Jan 15, 1988·Cell·R M Evans, S M Hollenberg
Jan 1, 1987·Methods in Enzymology·T A KunkelR A Zakour
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

Apr 2, 1992·Nature·R MarmorsteinS C Harrison
Jun 5, 1998·The Journal of Biological Chemistry·J C CortonS A Johnston
Jun 1, 1997·Fungal Genetics and Biology : FG & B·R B Todd, A Andrianopoulos
Dec 31, 1997·Fungal Genetics and Biology : FG & B·R B ToddM J Hynes

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