Sequential assignments of the sup 1 H NMR resonances of Zn(II) sub 2 and sup 113 Cd(II) sub 2 derivatives of the DNA-binding domain of the GAL4 transcription factor reveal a novel structural motif for specific DNA recognition

Journal Article · Tue Apr 30 00:00:00 EDT 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00231a016· OSTI ID:5394302

Pan, Tao; Coleman, J E ^[1]

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1. Yale Univ., New Haven, CT (USA)

The DNA-binding domain of the GAL4 transcription factor, consisting of the 62 N-terminal amino acid residues and denoted GAL4(62*), contains a novel Zn(II){sub 2}Cys{sub 6} or Cd(II){sub 2}Cys{sub 6} binuclear cluster. Specific DNA recognition requires residues located within as well as C terminal to this binuclear cluster. {sup 1}H NMR sequential assignments have been carried out on Zn(II){sub 2{minus}}- and {sup 113}Cd(II){sub 2}GAL4(62*) by using DQF-COSY, relayed COSY, double-relayed COSY, and NOESY. The ligands of the two tetrahedral metal-binding sites have been identified as Cys{sup 11}, Cys{sup 14}, Cys{sup 21}, and Cys{sup 31} to one metal ion and Cys{sup 28}, Cys{sup 38}, Cys{sup 21}, and Cys{sup 31} to the other metal ion with Cys{sup 21} and Cys{sup 31} as ligands shared between the two metal ions. No {alpha}-helices can be found within the GAL4(62*) structure, which consists of a series of turns to accommodate the metal cluster, followed by irregular loops and turns from residues 42 to 60, the specificity region, whose sequence contributes importantly to specific DNA recognition. Long-distance NOE's are observed between residues forming the binuclear cluster and several residues within the specificity region, indicating that the latter is folded compactly onto the metal cluster. The requirement of the Zn(II){sub 2}Cys{sub 6} binuclear cluster and the specificity region for binding to DNA reveals GAL4 as a member of a class of specific DNA-binding proteins using a new structural motif for the recognition of specific DNA sequences. Specific DNA binding by this class of proteins is achieved by use of turns and loops that enclose a Zn(II){sub 2}Cys{sub 6} binuclear cluster, instead of {alpha}-helices or {beta}-strands as observed in specific DNA-binding proteins described previously.

• OSTI ID: 5394302
• Journal Information: Biochemistry; (United States), Vol. 30:17; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
TRANSCRIPTION FACTORS
BIOCHEMICAL REACTION KINETICS
STRUCTURE-ACTIVITY RELATIONSHIPS
AMINO ACID SEQUENCE
CADMIUM 113
CADMIUM COMPOUNDS
MOLECULAR STRUCTURE
NUCLEAR MAGNETIC RESONANCE
OVERHAUSER EFFECT
SACCHAROMYCES CEREVISIAE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CADMIUM ISOTOPES
EUMYCOTA
EVEN-ODD NUCLEI
FUNGI
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
ISOMERIC TRANSITION ISOTOPES
ISOTOPES
KINETICS
MAGNETIC RESONANCE
MICROORGANISMS
NUCLEI
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PLANTS
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RESONANCE
SACCHAROMYCES
STABLE ISOTOPES
YEARS LIVING RADIOISOTOPES
YEASTS
550201* - Biochemistry- Tracer Techniques