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ID: 124494.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Inducing and modulating anisotropic DNA bends by pseudocomplementary peptide nucleic acids
Authors:Kuhn, H.; Cherny, D. I.; Demidov, V. V.; Frank-Kamenetskii, M. D.
Language:English
Date of Publication (YYYY-MM-DD):2004-05-18
Title of Journal:Proceedings of the National Academy of Sciences of the United States of America
Volume:101
Issue / Number:20
Start Page:7548
End Page:7553
Sequence Number of Article:doi:10.1073/pnas.0308756101
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:DNA bending is significant for various DNA functions in the cell. Here, we demonstrate that pseudocomplementary peptide nucleic acids (pcPNAs) represent a class of versatile, sequence-specific DNA-bending agents. The occurrence of anisotropic DNA bends induced by pcPNAs is shown by gel electrophoretic phasing analysis. The magnitude of DNA bending is determined by circular permutation assay and by electron microscopy, with good agreement of calculated mean values between both methods. Binding of a pair of 10-meric pcPNAs to its target DNA sequence results in moderate DNA bending with a mean value of 40–45°, while binding of one self-pc 8-mer PNA to target DNA yields a somewhat larger average value of the induced DNA bend. Both bends are found to be in phase when the pcPNA target sites are separated by distances of half-integer numbers of helical turns of regular duplex DNA, resulting in an enhanced DNA bend with an average value in the range of 80–90°. The occurrence of such a sharp bend within the DNA double helix is confirmed and exploited through efficient formation of 170-bp-long DNA minicircles by means of dimerization of two bent DNA fragments. The pcPNAs offer two main advantages over previously designed classes of nonnatural DNAbending agents: they have very mild sequence limitations while targeting duplex DNA and they can easily be designed for a chosen target sequence, because their binding obeys the principle of complementarity. We conclude that pcPNAs are promising tools for inducing bends in DNA at virtually any chosen site.
Last Change of the Resource (YYYY-MM-DD):2004-05-24
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Center for Advanced Biotechnology and Department of Biomedical Engineering, Boston University, 36 Cummington Street, Boston, MA 02215, USA; Institute of Molecular Genetics, Russian Academy of Sciences, Kurchatov's Square, Moscow, 123182, Russia
Identifiers:URL:http://www.pnas.org/content/101/20/7548.full
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