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Folding transition in the DNA-binding domain of GCN4 on specific binding to DNA.
Nature. 1990 Oct 11; 347(6293):575-8.Nat

Abstract

Protein-DNA recognition is often mediated by a small domain containing a recognizable structural motif, such as the helix-turn-helix or the zinc-finger. These motifs are compact structures that dock against the DNA double helix. Another DNA recognition motif, found in a highly conserved family of eukaryotic transcription factors including C/EPB, Fos, Jun and CREB, consists of a coiled-coil dimerization element the leucine-zipper and an adjoining basic region which mediates DNA binding. Here we describe circular dichroism and 1H-NMR spectroscopic studies of another family member, the yeast transcriptional activator GCN4. The 58-residue DNA-binding domain of GCN4, GCN4-p, exhibits a concentration-dependent alpha-helical transition, in accord with previous studies of the dimerization properties of an isolated leucine-zipper peptide. The GCN4-p dimer is approximately 70% helical at 25 degrees C, implying that the basic region adjacent to the leucine zipper is largely unstructured in the absence of DNA. Strikingly, addition of DNA containing a GCN4 binding site (AP-1 site) increases the alpha-helix content of GNC4-p to at least 95%. Thus, the basic region acquires substantial alpha-helical structure when it binds to DNA. A similar folding transition is observed on GCN4-p binding to the related ATF/CREB site, which contains an additional central base pair. The accommodation of DNA target sites of different lengths clearly requires some flexibility in the GCN4 binding domain, despite its high alpha-helix content. Our results indicate that the GCN4 basic region is significantly unfolded at 25 degrees C and that its folded, alpha-helical conformation is stabilized by binding to DNA.

Authors+Show Affiliations

Weiss MA
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
Ellenberger T
No affiliation info available
Wobbe CR
No affiliation info available
Lee JP
No affiliation info available
Harrison SC
No affiliation info available
Struhl K
No affiliation info available

MeSH

Activating Transcription FactorsAmino Acid SequenceBase SequenceBinding SitesBlood ProteinsCircular DichroismCyclic AMP Response Element-Binding ProteinDNADNA-Binding ProteinsFungal ProteinsLeucine ZippersMacromolecular SubstancesMagnetic Resonance SpectroscopyMolecular Sequence DataProtein ConformationProtein KinasesProto-Oncogene Proteins c-junSaccharomyces cerevisiae ProteinsTranscription Factors

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

2145515

Citation

Weiss, M A., et al. "Folding Transition in the DNA-binding Domain of GCN4 On Specific Binding to DNA." Nature, vol. 347, no. 6293, 1990, pp. 575-8.
Weiss MA, Ellenberger T, Wobbe CR, et al. Folding transition in the DNA-binding domain of GCN4 on specific binding to DNA. Nature. 1990;347(6293):575-8.
Weiss, M. A., Ellenberger, T., Wobbe, C. R., Lee, J. P., Harrison, S. C., & Struhl, K. (1990). Folding transition in the DNA-binding domain of GCN4 on specific binding to DNA. Nature, 347(6293), 575-8.
Weiss MA, et al. Folding Transition in the DNA-binding Domain of GCN4 On Specific Binding to DNA. Nature. 1990 Oct 11;347(6293):575-8. PubMed PMID: 2145515.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Folding transition in the DNA-binding domain of GCN4 on specific binding to DNA. AU - Weiss,M A, AU - Ellenberger,T, AU - Wobbe,C R, AU - Lee,J P, AU - Harrison,S C, AU - Struhl,K, PY - 1990/10/11/pubmed PY - 1990/10/11/medline PY - 1990/10/11/entrez SP - 575 EP - 8 JF - Nature JO - Nature VL - 347 IS - 6293 N2 - Protein-DNA recognition is often mediated by a small domain containing a recognizable structural motif, such as the helix-turn-helix or the zinc-finger. These motifs are compact structures that dock against the DNA double helix. Another DNA recognition motif, found in a highly conserved family of eukaryotic transcription factors including C/EPB, Fos, Jun and CREB, consists of a coiled-coil dimerization element the leucine-zipper and an adjoining basic region which mediates DNA binding. Here we describe circular dichroism and 1H-NMR spectroscopic studies of another family member, the yeast transcriptional activator GCN4. The 58-residue DNA-binding domain of GCN4, GCN4-p, exhibits a concentration-dependent alpha-helical transition, in accord with previous studies of the dimerization properties of an isolated leucine-zipper peptide. The GCN4-p dimer is approximately 70% helical at 25 degrees C, implying that the basic region adjacent to the leucine zipper is largely unstructured in the absence of DNA. Strikingly, addition of DNA containing a GCN4 binding site (AP-1 site) increases the alpha-helix content of GNC4-p to at least 95%. Thus, the basic region acquires substantial alpha-helical structure when it binds to DNA. A similar folding transition is observed on GCN4-p binding to the related ATF/CREB site, which contains an additional central base pair. The accommodation of DNA target sites of different lengths clearly requires some flexibility in the GCN4 binding domain, despite its high alpha-helix content. Our results indicate that the GCN4 basic region is significantly unfolded at 25 degrees C and that its folded, alpha-helical conformation is stabilized by binding to DNA. SN - 0028-0836 UR - https://www.unboundmedicine.com/medline/citation/2145515/Folding_transition_in_the_DNA_binding_domain_of_GCN4_on_specific_binding_to_DNA_ DB - PRIME DP - Unbound Medicine ER -