Solution structure of the Kluyveromyces lactis LAC9 Cd2 Cys6 DNA-binding domain.
The Zn2Cys6 DNA-binding domain has been identified by sequence homology in approximately forty fungal proteins, including the K. lactis LAC9 transcriptional activator. Using 1H NMR spectroscopy, we have determined the solution structure of a cadmium-substituted form of the LAC9 DNA-binding domain. We have complemented this approach by applying a series of 113Cd-1H NMR experiments, including several novel heteroTOCSY-based techniques. The DNA-binding domain forms a core of two alpha-helix/extended strand segments around the Cd2 binuclear cluster, with a network of amide proton-cysteinyl S gamma hydrogen bonds stabilizing the cluster. Comparison with other Zn2Cys6 domain structures provides insight into the common structural elements used in metal coordination and DNA binding.
Gardner
K H
KH
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114, USA.
Anderson
S F
SF
Coleman
J E
JE
D000595:Amino Acid Sequence; D001665:Binding Sites; D002104:Cadmium / Q000737:chemistry* / Q000378:metabolism; D003545:Cysteine; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000737:chemistry*; D007716:Kluyveromyces / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C058596:LAC9 protein, Kluyveromyces lactis; D014157:Transcription Factors; D002104:Cadmium; D004247:DNA; D003545:Cysteine