Hybridization of G-quadruplex forming peptide nucleic acids to guanine rich DNA templates inhibits DNA polymerase η extension.

Hybridization of G-quadruplex forming peptide nucleic acids to guanine rich DNA templates inhibits DNA polymerase η extension. Murphy, C. T, Gupta, A., Armitage, B. A., & Opresko, P. L Biochemistry, July, 2014. ISBN: 4126248285

Paper doi abstract bibtex

The guanine quadruplex (G-quadruplex) is a highly stable secondary structure that forms in G-rich repeats of DNA, which can interfere with DNA processes including DNA replication and transcription. We showed previously that short guanine rich Peptide Nucleic Acids (PNAs) can form highly stable hybrid quadruplexes with DNA. We hypothesized that such structures would provide a stronger block to polymerase extension on G-rich templates than a native DNA homoquadruplex due to the greater thermodynamic stability of the PNA/DNA hybrid structures. To test this we analyzed DNA primer extension activity of polymerase η, a translesion polymerase implicated in synthesis past G-quadruplex blocks, on DNA templates containing guanine repeats. We observed a PNA concentration dependent decrease in polymerase η extension to the end of the template, and an increase in polymerase η inhibition at the sequence prior to the G-rich repeats. In contrast, the addition of a complementary C-rich PNA that hybridizes to the G-rich repeats by Watson-Crick base pairing led to a decrease in polymerase inhibition and an increase in full length extension products. The G-quadruplex forming PNA exhibited greater inhibition (IC50 = 16.2 nM) of polymerase η DNA synthesis on the G-rich templates when compared to the established G-quadruplex stabilizing ligand BRACO-19 (IC50 = 42.5 nM). Our results indicate that homologous PNA targeting of G-rich sequences creates stable PNA/DNA heteroquadruplexes that inhibit polymerase η extension more effectively than a DNA homoquadruplex. The implications of these results for potential development of homologous PNAs as therapeutics for halting proliferating cancer cells are discussed.

@article{Murphy2014,
	title = {Hybridization of {G}-quadruplex forming peptide nucleic acids to guanine rich {DNA} templates inhibits {DNA} polymerase η extension.},
	issn = {1520-4995},
	url = {http://www.ncbi.nlm.nih.gov/pubmed/25068499},
	doi = {10.1021/bi5006859},
	abstract = {The guanine quadruplex (G-quadruplex) is a highly stable secondary structure that forms in G-rich repeats of DNA, which can interfere with DNA processes including DNA replication and transcription. We showed previously that short guanine rich Peptide Nucleic Acids (PNAs) can form highly stable hybrid quadruplexes with DNA. We hypothesized that such structures would provide a stronger block to polymerase extension on G-rich templates than a native DNA homoquadruplex due to the greater thermodynamic stability of the PNA/DNA hybrid structures. To test this we analyzed DNA primer extension activity of polymerase η, a translesion polymerase implicated in synthesis past G-quadruplex blocks, on DNA templates containing guanine repeats. We observed a PNA concentration dependent decrease in polymerase η extension to the end of the template, and an increase in polymerase η inhibition at the sequence prior to the G-rich repeats. In contrast, the addition of a complementary C-rich PNA that hybridizes to the G-rich repeats by Watson-Crick base pairing led to a decrease in polymerase inhibition and an increase in full length extension products. The G-quadruplex forming PNA exhibited greater inhibition (IC50 = 16.2 nM) of polymerase η DNA synthesis on the G-rich templates when compared to the established G-quadruplex stabilizing ligand BRACO-19 (IC50 = 42.5 nM). Our results indicate that homologous PNA targeting of G-rich sequences creates stable PNA/DNA heteroquadruplexes that inhibit polymerase η extension more effectively than a DNA homoquadruplex. The implications of these results for potential development of homologous PNAs as therapeutics for halting proliferating cancer cells are discussed.},
	journal = {Biochemistry},
	author = {Murphy, Connor T and Gupta, Anisha and Armitage, Bruce Alan and Opresko, Patricia L},
	month = jul,
	year = {2014},
	pmid = {25068499},
	note = {ISBN: 4126248285},
	keywords = {\#nosource},
}

Downloads: 0

{"_id":"d5zn4e3XX8TyFnve7","bibbaseid":"murphy-gupta-armitage-opresko-hybridizationofgquadruplexformingpeptidenucleicacidstoguaninerichdnatemplatesinhibitsdnapolymeraseextension-2014","author_short":["Murphy, C. T","Gupta, A.","Armitage, B. A.","Opresko, P. L"],"bibdata":{"bibtype":"article","type":"article","title":"Hybridization of G-quadruplex forming peptide nucleic acids to guanine rich DNA templates inhibits DNA polymerase η extension.","issn":"1520-4995","url":"http://www.ncbi.nlm.nih.gov/pubmed/25068499","doi":"10.1021/bi5006859","abstract":"The guanine quadruplex (G-quadruplex) is a highly stable secondary structure that forms in G-rich repeats of DNA, which can interfere with DNA processes including DNA replication and transcription. We showed previously that short guanine rich Peptide Nucleic Acids (PNAs) can form highly stable hybrid quadruplexes with DNA. We hypothesized that such structures would provide a stronger block to polymerase extension on G-rich templates than a native DNA homoquadruplex due to the greater thermodynamic stability of the PNA/DNA hybrid structures. To test this we analyzed DNA primer extension activity of polymerase η, a translesion polymerase implicated in synthesis past G-quadruplex blocks, on DNA templates containing guanine repeats. We observed a PNA concentration dependent decrease in polymerase η extension to the end of the template, and an increase in polymerase η inhibition at the sequence prior to the G-rich repeats. In contrast, the addition of a complementary C-rich PNA that hybridizes to the G-rich repeats by Watson-Crick base pairing led to a decrease in polymerase inhibition and an increase in full length extension products. The G-quadruplex forming PNA exhibited greater inhibition (IC50 = 16.2 nM) of polymerase η DNA synthesis on the G-rich templates when compared to the established G-quadruplex stabilizing ligand BRACO-19 (IC50 = 42.5 nM). Our results indicate that homologous PNA targeting of G-rich sequences creates stable PNA/DNA heteroquadruplexes that inhibit polymerase η extension more effectively than a DNA homoquadruplex. The implications of these results for potential development of homologous PNAs as therapeutics for halting proliferating cancer cells are discussed.","journal":"Biochemistry","author":[{"propositions":[],"lastnames":["Murphy"],"firstnames":["Connor","T"],"suffixes":[]},{"propositions":[],"lastnames":["Gupta"],"firstnames":["Anisha"],"suffixes":[]},{"propositions":[],"lastnames":["Armitage"],"firstnames":["Bruce","Alan"],"suffixes":[]},{"propositions":[],"lastnames":["Opresko"],"firstnames":["Patricia","L"],"suffixes":[]}],"month":"July","year":"2014","pmid":"25068499","note":"ISBN: 4126248285","keywords":"#nosource","bibtex":"@article{Murphy2014,\n\ttitle = {Hybridization of {G}-quadruplex forming peptide nucleic acids to guanine rich {DNA} templates inhibits {DNA} polymerase η extension.},\n\tissn = {1520-4995},\n\turl = {http://www.ncbi.nlm.nih.gov/pubmed/25068499},\n\tdoi = {10.1021/bi5006859},\n\tabstract = {The guanine quadruplex (G-quadruplex) is a highly stable secondary structure that forms in G-rich repeats of DNA, which can interfere with DNA processes including DNA replication and transcription. We showed previously that short guanine rich Peptide Nucleic Acids (PNAs) can form highly stable hybrid quadruplexes with DNA. We hypothesized that such structures would provide a stronger block to polymerase extension on G-rich templates than a native DNA homoquadruplex due to the greater thermodynamic stability of the PNA/DNA hybrid structures. To test this we analyzed DNA primer extension activity of polymerase η, a translesion polymerase implicated in synthesis past G-quadruplex blocks, on DNA templates containing guanine repeats. We observed a PNA concentration dependent decrease in polymerase η extension to the end of the template, and an increase in polymerase η inhibition at the sequence prior to the G-rich repeats. In contrast, the addition of a complementary C-rich PNA that hybridizes to the G-rich repeats by Watson-Crick base pairing led to a decrease in polymerase inhibition and an increase in full length extension products. The G-quadruplex forming PNA exhibited greater inhibition (IC50 = 16.2 nM) of polymerase η DNA synthesis on the G-rich templates when compared to the established G-quadruplex stabilizing ligand BRACO-19 (IC50 = 42.5 nM). Our results indicate that homologous PNA targeting of G-rich sequences creates stable PNA/DNA heteroquadruplexes that inhibit polymerase η extension more effectively than a DNA homoquadruplex. The implications of these results for potential development of homologous PNAs as therapeutics for halting proliferating cancer cells are discussed.},\n\tjournal = {Biochemistry},\n\tauthor = {Murphy, Connor T and Gupta, Anisha and Armitage, Bruce Alan and Opresko, Patricia L},\n\tmonth = jul,\n\tyear = {2014},\n\tpmid = {25068499},\n\tnote = {ISBN: 4126248285},\n\tkeywords = {\\#nosource},\n}\n\n","author_short":["Murphy, C. T","Gupta, A.","Armitage, B. A.","Opresko, P. L"],"key":"Murphy2014","id":"Murphy2014","bibbaseid":"murphy-gupta-armitage-opresko-hybridizationofgquadruplexformingpeptidenucleicacidstoguaninerichdnatemplatesinhibitsdnapolymeraseextension-2014","role":"author","urls":{"Paper":"http://www.ncbi.nlm.nih.gov/pubmed/25068499"},"keyword":["#nosource"],"metadata":{"authorlinks":{}},"html":""},"bibtype":"article","biburl":"https://bibbase.org/zotero/eric.larG4","dataSources":["4i5C7S78DvJNsaHyg","5L2zM5wNE5CBYNuea"],"keywords":["#nosource"],"search_terms":["hybridization","quadruplex","forming","peptide","nucleic","acids","guanine","rich","dna","templates","inhibits","dna","polymerase","extension","murphy","gupta","armitage","opresko"],"title":"Hybridization of G-quadruplex forming peptide nucleic acids to guanine rich DNA templates inhibits DNA polymerase η extension.","year":2014}