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Evidence that a minor groove-binding peptide and a major groove-binding protein can simultaneously occupy a common site on DNA

Oakley, Martha G. and Mrksich, Milan and Dervan, Peter B. (1992) Evidence that a minor groove-binding peptide and a major groove-binding protein can simultaneously occupy a common site on DNA. Biochemistry, 31 (45). pp. 10969-10975. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20160510-142114334

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Abstract

Affinity cleaving proteins have been synthesized based on the DNA-binding domain of the yeast transcriptional activator GCN4 with the DNA cleaving moiety Fe·EDTA attached at the NH_2 terminus [Oakley, M. G., & Dervan, P. B. (1990) Science 248, 847]. Cleavage patterns generated by Fe·EDTA-GCN4(226-281) bound to the DNA sites 5'-CTGACTAAT-3' and 5'-ATGACTCTT-3' reveal that the NH_2 termini of the GCN4 DNA-binding domain are located in the major groove of DNA, 9-10 base pairs apart, consistent with a Y-shaped dimeric structure. 1-Methylimidazole-2-carboxamide netropsin (2-ImN) is a designed synthetic peptide which binds in the minor groove of DNA at 5'-TGACT-3' sites as an anti parallel, side-by-side dimer [Mrksich, M., Wade, W. S., Dwyer, T. J ., Geierstanger, B. H., Wemmer, D. E., & Dervan, P. B. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 7586]. Through the use of Fe·EDTAGCN4( 226-281) as a sequence-specific footprinting agent, it is shown that the dimeric protein GCN4-(226-281) and the dimeric peptide 2-ImN can simultaneously occupy their common binding site in the major and minor grooves of DNA, respectively. The association constants for 2-ImN in the presence and in the absence of Fe·EDTA-GCN4(226-281) are found to be similar, suggesting that the binding of the two dimers is not cooperative.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bi00160a005DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi00160a005PublisherArticle
ORCID:
AuthorORCID
Dervan, Peter B.0000-0001-8852-7306
Additional Information:© 1992 American Chemical Society. Received May 18, 1992; Revised Manuscript Received August 24, 1992. We thank C. S. Parker for the gift of the plasmid pARE/GCRE, S. R. Wilson (New York University) for electrospray mass spectral analysis, and S. F. Singleton and W. S. Wade for helpful discussions. This work was supported by the National Institutes of Health (GM-27681), the National Foundation for Cancer Research, a National Science Foundation predoctoral fellowship, and a National Institutes of Health predoctoral traineeship to M.G.O. and a National Institutes of Health Research Service Award to M.M.
Funders:
Funding AgencyGrant Number
NIHGM-27681
National Foundation for Cancer ResearchUNSPECIFIED
NSF Predoctoral FellowshipUNSPECIFIED
NIH Predoctoral FellowshipUNSPECIFIED
Issue or Number:45
Record Number:CaltechAUTHORS:20160510-142114334
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160510-142114334
Official Citation:Evidence that a minor groove-binding peptide and a major groove-binding protein can simultaneously occupy a common site on DNA Martha G. Oakley, Milan Mrksich, and Peter B. Dervan Biochemistry 1992 31 (45), 10969-10975 DOI: 10.1021/bi00160a005
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:66941
Collection:CaltechAUTHORS
Deposited By: Victoria Brennan
Deposited On:19 May 2016 18:13
Last Modified:26 Nov 2019 11:15

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