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Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs
Numerous regulatory genes have G-rich regions that can potentially form quadruplex structures, possibly playing a role in transcription regulation. We studied a G-rich sequence in the BCL2 gene 176-bp upstream of the P1 promoter for G-quadruplex formation. Using circular dichroism (CD), thermal dena...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2794188/ https://www.ncbi.nlm.nih.gov/pubmed/19820116 http://dx.doi.org/10.1093/nar/gkp840 |
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author | Onyshchenko, Mykola I. Gaynutdinov, Timur I. Englund, Ethan A. Appella, Daniel H. Neumann, Ronald D. Panyutin, Igor G. |
author_facet | Onyshchenko, Mykola I. Gaynutdinov, Timur I. Englund, Ethan A. Appella, Daniel H. Neumann, Ronald D. Panyutin, Igor G. |
author_sort | Onyshchenko, Mykola I. |
collection | PubMed |
description | Numerous regulatory genes have G-rich regions that can potentially form quadruplex structures, possibly playing a role in transcription regulation. We studied a G-rich sequence in the BCL2 gene 176-bp upstream of the P1 promoter for G-quadruplex formation. Using circular dichroism (CD), thermal denaturation and dimethyl sulfate (DMS) footprinting, we found that a single-stranded oligonucleotide with the sequence of the BCL2 G-rich region forms a potassium-stabilized G-quadruplex. To study G-quadruplex formation in double-stranded DNA, the G-rich sequence of the BCL2 gene was inserted into plasmid DNA. We found that a G-quadruplex did not form in the insert at physiological conditions. To induce G-quadruplex formation, we used short peptide nucleic acids (PNAs) that bind to the complementary C-rich strand. We examined both short duplex-forming PNAs, complementary to the central part of the BCL2 gene, and triplex-forming bis-PNAs, complementary to sequences adjacent to the G-rich BCL2 region. Using a DMS protection assay, we demonstrated G-quadruplex formation within the G-rich sequence from the promoter region of the human BCL2 gene in plasmid DNA. Our results show that molecules binding the complementary C-strand facilitate G-quadruplex formation and introduce a new mode of PNA-mediated sequence-specific targeting. |
format | Text |
id | pubmed-2794188 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-27941882009-12-16 Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs Onyshchenko, Mykola I. Gaynutdinov, Timur I. Englund, Ethan A. Appella, Daniel H. Neumann, Ronald D. Panyutin, Igor G. Nucleic Acids Res Molecular Biology Numerous regulatory genes have G-rich regions that can potentially form quadruplex structures, possibly playing a role in transcription regulation. We studied a G-rich sequence in the BCL2 gene 176-bp upstream of the P1 promoter for G-quadruplex formation. Using circular dichroism (CD), thermal denaturation and dimethyl sulfate (DMS) footprinting, we found that a single-stranded oligonucleotide with the sequence of the BCL2 G-rich region forms a potassium-stabilized G-quadruplex. To study G-quadruplex formation in double-stranded DNA, the G-rich sequence of the BCL2 gene was inserted into plasmid DNA. We found that a G-quadruplex did not form in the insert at physiological conditions. To induce G-quadruplex formation, we used short peptide nucleic acids (PNAs) that bind to the complementary C-rich strand. We examined both short duplex-forming PNAs, complementary to the central part of the BCL2 gene, and triplex-forming bis-PNAs, complementary to sequences adjacent to the G-rich BCL2 region. Using a DMS protection assay, we demonstrated G-quadruplex formation within the G-rich sequence from the promoter region of the human BCL2 gene in plasmid DNA. Our results show that molecules binding the complementary C-strand facilitate G-quadruplex formation and introduce a new mode of PNA-mediated sequence-specific targeting. Oxford University Press 2009-12 2009-10-09 /pmc/articles/PMC2794188/ /pubmed/19820116 http://dx.doi.org/10.1093/nar/gkp840 Text en © The Author(s) 2009. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Molecular Biology Onyshchenko, Mykola I. Gaynutdinov, Timur I. Englund, Ethan A. Appella, Daniel H. Neumann, Ronald D. Panyutin, Igor G. Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title | Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title_full | Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title_fullStr | Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title_full_unstemmed | Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title_short | Stabilization of G-quadruplex in the BCL2 promoter region in double-stranded DNA by invading short PNAs |
title_sort | stabilization of g-quadruplex in the bcl2 promoter region in double-stranded dna by invading short pnas |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2794188/ https://www.ncbi.nlm.nih.gov/pubmed/19820116 http://dx.doi.org/10.1093/nar/gkp840 |
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