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Title: 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

Abstract

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 ofmore » 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.« less

Authors:
;  [1]
  1. Yale Univ., New Haven, CT (USA)
Publication Date:
OSTI Identifier:
5394302
Resource Type:
Journal Article
Journal Name:
Biochemistry; (United States)
Additional Journal Information:
Journal Volume: 30:17; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Pan, Tao, and Coleman, J E. 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. United States: N. p., 1991. Web. doi:10.1021/bi00231a016.
Pan, Tao, & Coleman, J E. 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. United States. https://doi.org/10.1021/bi00231a016
Pan, Tao, and Coleman, J E. 1991. "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". United States. https://doi.org/10.1021/bi00231a016.
@article{osti_5394302,
title = {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},
author = {Pan, Tao and Coleman, J E},
abstractNote = {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.},
doi = {10.1021/bi00231a016},
url = {https://www.osti.gov/biblio/5394302}, journal = {Biochemistry; (United States)},
issn = {0006-2960},
number = ,
volume = 30:17,
place = {United States},
year = {Tue Apr 30 00:00:00 EDT 1991},
month = {Tue Apr 30 00:00:00 EDT 1991}
}