Circular Dichroism Spectra and Electrophoretic Mobility Shift Assays Show That Human Replication Protein A Binds and Melts Intramolecular G-Quadruplex Structures†,‡

Biochemistry ◽  
2009 ◽  
Vol 48 (5) ◽  
pp. 1099-1111 ◽  
Author(s):  
Jun-Huei Fan ◽  
Elena Bochkareva ◽  
Alexey Bochkarev ◽  
Donald M. Gray
Keyword(s):  
Circular Dichroism ◽  
Electrophoretic Mobility ◽  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Circular Dichroism Spectra ◽  
Mobility Shift ◽  
G Quadruplex ◽  
Electrophoretic Mobility Shift Assays ◽  
Circular Dichroïsm

scholarly journals Stabilization of a G-Quadruplex from Unfolding by Replication Protein A Using Potassium and the Porphyrin TMPyP4

2011 ◽  
Vol 2011 ◽  
pp. 1-13 ◽  
Author(s):  
Aishwarya Prakash ◽  
Fabien Kieken ◽  
Luis A. Marky ◽  
Gloria E. O. Borgstahl
Keyword(s):  
Dna Replication ◽  
Protein A ◽  
Thermal Stabilization ◽  
Replication Protein A ◽  
Replication Protein ◽  
Unique Problem ◽  
Ssdna Binding ◽  
Binding Domains ◽  
G Quadruplex ◽  
Stabilization Effect

Replication protein A (RPA) plays an essential role in DNA replication by binding and unfolding non-canonical single-stranded DNA (ssDNA) structures. Of the six RPA ssDNA binding domains (labeled A-F), RPA-CDE selectively binds a G-quadruplex forming sequence (5′-TAGGGGAAGGGTTGGAGTGGGTT-3′called Gq23). In K+, Gq23 forms a mixed parallel/antiparallel conformation, and in Na+Gq23 has a less stable (TMlowered by ∼20∘C), antiparallel conformation. Gq23 is intramolecular and 1D NMR confirms a stable G-quadruplex structure in K+. Full-length RPA and RPA-CDE-core can bind and unfold the Na+form of Gq23 very efficiently, but complete unfolding is not observed with the K+form. Studies with G-quadruplex ligands, indicate that TMPyP4 has a thermal stabilization effect on Gq23 in K+, and inhibits complete unfolding by RPA and RPA-CDE-core. Overall these data indicate that G-quadruplexes present a unique problem for RPA to unfold and ligands, such as TMPyP4, could possibly hinder DNA replication by blocking unfolding by RPA.

scholarly journals Analysis of a meiosis-specific URS1 site: sequence requirements and involvement of replication protein A.

1997 ◽  
Vol 17 (7) ◽  
pp. 3536-3546 ◽  
Author(s):  
V Gailus-Durner ◽  
C Chintamaneni ◽  
R Wilson ◽  
S J Brill ◽  
A K Vershon
Keyword(s):  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Sequence Specificity ◽  
Mobility Shift ◽  
Cell Extracts ◽  
Yeast Genes ◽  
Sequence Requirements

URS1 is a transcriptional repressor site found in the promoters of a wide variety of yeast genes that are induced under stress conditions. In the context of meiotic promoters, URS1 sites act as repressor sequences during mitosis and function as activator sites during meiosis. We have investigated the sequence requirements of the URS1 site of the meiosis-specific HOP1 gene (URS1H) and have found differences compared with a URS1 site from a nonmeiotic gene. We have also observed that the sequence specificity for meiotic activation at this site differs from that for mitotic repression. Base pairs flanking the conserved core sequence enhance meiotic induction but are not required for mitotic repression of HOP1. Electrophoretic mobility shift assays of mitotic and meiotic cell extracts show a complex pattern of DNA-protein complexes, suggesting that several different protein factors bind specifically to the site. We have determined that one of the complexes of URS1H is formed by replication protein A (RPA). Although RPA binds to the double-stranded URS1H site in vitro, it has much higher affinity for single-stranded than for double-stranded URS1H, and one-hybrid assays suggest that RPA does not bind to this site at detectable levels in vivo. In addition, conditional-lethal mutations in RPA were found to have no effect on URS1H-mediated repression. These results suggest that although RPA binds to URS1H in vitro, it does not appear to have a functional role in transcriptional repression through this site in vivo.

Interaction of long telomeric DNAs with macrocyclic hexaoxazole as a G-quadruplex ligand

MedChemComm ◽  
2013 ◽  
Vol 4 (1) ◽  
pp. 260-264 ◽  
Author(s):  
Keisuke Iida ◽  
Gen Tsubouchi ◽  
Takahiro Nakamura ◽  
Satoki Majima ◽  
Hiroyuki Seimiya ◽  
...  
Keyword(s):  
Circular Dichroism ◽  
Living Cells ◽  
Dna Melting ◽  
Mobility Shift ◽  
Electrophoresis Mobility Shift Assay ◽  
G Quadruplex ◽  
Mobility Shift Assay ◽  
Circular Dichroïsm

The interactions of long telomeric DNAs, which mimic telomeres in living cells, with a macrocyclic hexaoxazole ligand L2H2-6OTD (2) were investigated by means of electrophoresis mobility shift assay, circular dichroism (CD) titration analysis, and DNA melting measurements.

scholarly journals Interaction of Replication Protein-A with G-Quadruplex Structures

2012 ◽  
Vol 102 (3) ◽  
pp. 71a
Author(s):  
Sujay Ray ◽  
Mohammad Haroon Qureshi ◽  
Dominic Malcom ◽  
Ugur Celik ◽  
Hamza Balci
Keyword(s):  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
G Quadruplex

scholarly journals Replication protein A plays multifaceted roles complementary to specialized helicases in processing G-quadruplex DNA

iScience ◽  
2021 ◽  
pp. 102493
Author(s):  
Yi-Ran Wang ◽  
Ting-Ting Guo ◽  
Ya-Ting Zheng ◽  
Chang-Wei Lai ◽  
Bo Sun ◽  
...  
Keyword(s):  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
Quadruplex Dna ◽  
G Quadruplex

scholarly journals Replication Protein A Unfolds G-Quadruplex Structures with Varying Degrees of Efficiency

2012 ◽  
Vol 116 (19) ◽  
pp. 5588-5594 ◽  
Author(s):  
Mohammad H. Qureshi ◽  
Sujay Ray ◽  
Abby L. Sewell ◽  
Soumitra Basu ◽  
Hamza Balci
Keyword(s):  
Protein A ◽  
Replication Protein A ◽  
Replication Protein ◽  
G Quadruplex
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