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Site-Specific Inhibition of Transcription Factor Binding to DNA by a Metallointercalator

Odom, Duncan T. and Parker, Carl S. and Barton, Jacqueline K. (1999) Site-Specific Inhibition of Transcription Factor Binding to DNA by a Metallointercalator. Biochemistry, 38 (16). pp. 5155-5163. ISSN 0006-2960. http://resolver.caltech.edu/CaltechAUTHORS:20160407-115524241

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Abstract

The metallointercalator Λ-1-Rh(MGP)_2phi^(5+) binds tightly and specifically to the site 5‘-CATATG-3‘ in the major groove of double helical DNA by a combination of direct readout and shape selection. To examine competitive interactions between this small metal complex and a DNA-binding transcription factor, the preferred binding site for Λ-1-Rh(MGP)_2phi^(5+) was engineered into the AP-1 recognition element (ARE) of the major-groove binding bZIP transcription factor yAP-1, the yeast analogue of mammalian AP-1. Binding experiments confirmed that the modified ARE retained normal yAP-1 binding affinity. Photocleavage experiments demonstrated that the modified ARE contained a high-affinity binding site for Λ-1-Rh(MGP)_2phi^(5+), whereas the native ARE showed no interaction. Competition experiments using gel shift mobility assays demonstrated that Λ-1-Rh(MGP)_2phi^(5+) at 120 nM competes 50% of yAP-1 binding to the 5‘-CATATG-3‘ containing oligonucleotide. In contrast, competitive disruption of protein binding to the native ARE requires 3 μM Λ-1-Rh(MGP)_2phi^(5+). Metallointercalator derivatives, including geometric isomers of Λ-1-Rh(MGP)_2phi^(5+), show no specific binding to the target site and show no inhibition of yAP-1/DNA complexes at concentrations as high as 20 μM. Thus, metallointercalators can be tuned to show selectivity for major groove sites on DNA comparable to transcription factors and indeed can inhibit transcription factor binding site selectively.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bi9827969DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi9827969PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/bi9827969Related ItemSupporting Information
ORCID:
AuthorORCID
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 1999 American Chemical Society. Received November 25, 1998; Revised Manuscript Received February 3, 1999. Publication Date (Web): April 1, 1999. We are grateful to the NIH (GM33309 to J.K.B. and GM47381 to C.S.P.) for their financial support and for an NRSA traineeship to D.T.O. We also thank Dr. Christine Rener for technical assistance.
Funders:
Funding AgencyGrant Number
NIHGM33309
NIHGM47381
NIH Predoctoral FellowshipUNSPECIFIED
Record Number:CaltechAUTHORS:20160407-115524241
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160407-115524241
Official Citation:Site-Specific Inhibition of Transcription Factor Binding to DNA by a Metallointercalator Duncan T. Odom, Carl S. Parker, and Jacqueline K. Barton Biochemistry 1999 38 (16), 5155-5163 DOI: 10.1021/bi9827969
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65992
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:11 Apr 2016 17:21
Last Modified:12 Apr 2016 22:07

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