| Autor: |
Eli Rothenberg, Dipika Gupta, Wei Ting C. Lee |
| Rok vydání: |
2021 |
| Předmět: |
|
| Zdroj: |
Methods in Enzymology ISBN: 9780323907330 |
| DOI: |
10.1016/bs.mie.2021.08.008 |
| Popis: |
DNA G-quadruplexes (G4s) are stable, non-canonical DNA secondary structures formed within guanine(G)-rich sequences. While extensively studied in vitro, evidence of the occurrence of G4s in vivo has only recently emerged. The formation of G4 structures may pose an obstacle for diverse DNA transactions including replication, which is linked to mutagenesis and genomic instability. A fundamental question in the field has been whether and how the formation of G4s is coupled to the progression of replication forks. This process has remained undefined largely due to the lack of experimental approaches capable of monitoring the presence of G4s and their association with the replication machinery in cells. Here, we describe a detailed multicolor single-molecule localization microscopy (SMLM) protocol for detecting nanoscale spatial-association of DNA G4s with the cellular replisome complex. This method offers a unique platform for visualizing the mechanisms of G4 formation at the molecular level, as well as addressing key biological questions as to the functional roles of these structures in the maintenance of genome integrity. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
