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Assembly of a G-Quadruplex Repair Complex by the FANCJ DNA Helicase and the REV1 Polymerase.

Genes (Basel). 2019; 
Lowran K1, Campbell L1, Popp P2, Wu CG1.
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Custom DNA/RNA Oligos … Experiments were performed at 25 °C in Buffer H (20 mM HEPES pH 7.5, 150 mM KCl, 5 mM<br> TCEP, and 5% (v/v) glycerol) unless otherwise specified. 2.2. FANCJ Peptides and DNA Oligos.<br> FANCJ peptides were synthesized by <b>Genscript</b> (Piscataway, NJ, USA) … Get A Quote

摘要

The FANCJ helicase unfolds G-quadruplexes (G4s) in human cells to support DNA replication. This action is coupled to the recruitment of REV1 polymerase to synthesize DNA across from a guanine template. The precise mechanisms of these reactions remain unclear. While FANCJ binds to G4s with an AKKQ motif, it is not known whether this site recognizes damaged G4 structures. FANCJ also has a PIP-like (PCNA Interacting Protein) region that may recruit REV1 to G4s either directly or through interactions mediated by PCNA protein. In this work, we measured the affinities of a FANCJ AKKQ peptide for G4s formed by (TTAGGG)4 and (GGGT)4 using fluorescence spectroscopy and biolayer interferometry (BLI). The effects of 8-oxo... More

关键词

8-oxoguanine (8oxoG); DNA repair; FANCJ; G-quadruplexes (G4s); PCNA; REV1; biolayer interferometry; circular dichroism; fluorescence spectroscopy; helicases