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G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA.

Publication ,  Journal Article
Giri, B; Smaldino, PJ; Thys, RG; Creacy, SD; Routh, ED; Hantgan, RR; Lattmann, S; Nagamine, Y; Akman, SA; Vaughn, JP
Published in: Nucleic Acids Res
September 1, 2011

It has been previously shown that the DHX36 gene product, G4R1/RHAU, tightly binds tetramolecular G4-DNA with high affinity and resolves these structures into single strands. Here, we test the ability of G4R1/RHAU to bind and unwind unimolecular G4-DNA. Gel mobility shift assays were used to measure the binding affinity of G4R1/RHAU for unimolecular G4-DNA-formed sequences from the Zic1 gene and the c-Myc promoter. Extremely tight binding produced apparent K(d)'s of 6, 3 and 4 pM for two Zic1 G4-DNAs and a c-Myc G4-DNA, respectively. The low enzyme concentrations required for measuring these K(d)'s limit the precision of their determination to upper boundary estimates. Similar tight binding was not observed in control non-G4 forming DNA sequences or in single-stranded DNA having guanine-rich runs capable of forming tetramolecular G4-DNA. Using a peptide nucleic acid (PNA) trap assay, we show that G4R1/RHAU catalyzes unwinding of unimolecular Zic1 G4-DNA into an unstructured state capable of hybridizing to a complementary PNA. Binding was independent of adenosine triphosphate (ATP), but the PNA trap assay showed that unwinding of G4-DNA was ATP dependent. Competition studies indicated that unimolecular Zic1 and c-Myc G4-DNA structures inhibit G4R1/RHAU-catalyzed resolution of tetramolecular G4-DNA. This report provides evidence that G4R1/RHAU tightly binds and unwinds unimolecular G4-DNA structures.

Published In

Nucleic Acids Res

DOI

EISSN

1362-4962

Publication Date

September 1, 2011

Volume

39

Issue

16

Start / End Page

7161 / 7178

Location

England

Related Subject Headings

  • Recombinases
  • Peptide Nucleic Acids
  • Oligodeoxyribonucleotides
  • Nucleic Acid Hybridization
  • Humans
  • Genes, myc
  • G-Quadruplexes
  • Developmental Biology
  • DNA
  • DEAD-box RNA Helicases
 

Citation

APA
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MLA
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Giri, B., Smaldino, P. J., Thys, R. G., Creacy, S. D., Routh, E. D., Hantgan, R. R., … Vaughn, J. P. (2011). G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA. Nucleic Acids Res, 39(16), 7161–7178. https://doi.org/10.1093/nar/gkr234
Giri, Banabihari, Philip J. Smaldino, Ryan G. Thys, Steven D. Creacy, Eric D. Routh, Roy R. Hantgan, Simon Lattmann, Yoshikuni Nagamine, Steven A. Akman, and James P. Vaughn. “G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA.Nucleic Acids Res 39, no. 16 (September 1, 2011): 7161–78. https://doi.org/10.1093/nar/gkr234.
Giri B, Smaldino PJ, Thys RG, Creacy SD, Routh ED, Hantgan RR, et al. G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA. Nucleic Acids Res. 2011 Sep 1;39(16):7161–78.
Giri, Banabihari, et al. “G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA.Nucleic Acids Res, vol. 39, no. 16, Sept. 2011, pp. 7161–78. Pubmed, doi:10.1093/nar/gkr234.
Giri B, Smaldino PJ, Thys RG, Creacy SD, Routh ED, Hantgan RR, Lattmann S, Nagamine Y, Akman SA, Vaughn JP. G4 resolvase 1 tightly binds and unwinds unimolecular G4-DNA. Nucleic Acids Res. 2011 Sep 1;39(16):7161–7178.
Journal cover image

Published In

Nucleic Acids Res

DOI

EISSN

1362-4962

Publication Date

September 1, 2011

Volume

39

Issue

16

Start / End Page

7161 / 7178

Location

England

Related Subject Headings

  • Recombinases
  • Peptide Nucleic Acids
  • Oligodeoxyribonucleotides
  • Nucleic Acid Hybridization
  • Humans
  • Genes, myc
  • G-Quadruplexes
  • Developmental Biology
  • DNA
  • DEAD-box RNA Helicases