DNA Synthesis | Dna Replication | Telomere
DNA Replication 3-D Animation from DNA ..
Chastain PD, Makhov AM 2nd, Nossal NG and Griffith J (2003) Architecture of the replication complex and DNA loops at the fork generated by the bacteriophage t4 proteins. Journal of Biological Chemistry 278(23): 21276–21285.
The DNA near each replication fork is coated with single-stranded ..
A comprehensive proteomics analysis of proteins at normal and stressed replication forks illuminates the role of ATR in preventing fork collapse.
where separation of the DNA occurs is called the replication fork
The key to iPOND is the use of “click” chemistry, a copper-catalyzed reaction between an azide and an alkyne, that has been employed extensively to label and retrieve biomolecules from the complex cellular environment (Figure 2). The Cortez lab took advantage of the fact that cells readily incorporate the alkyne-containing nucleotide 5′-ethynyl-2′-deoxyuridine (EdU) in place of thymidine as they replicate DNA. Reaction of DNA containing EdU with a biotin azide reagent (designed and synthesized in the lab of VICB member Ned Porter) under click chemistry conditions attaches a biotin label to the alkyne moiety of the incorporated EdU in the DNA. Following fragmentation of the DNA, biotin-labeled DNA fragments are captured on streptavidin-coated beads, providing a rapid and efficient method for purification (Figure 3A). The use of formaldehyde to covalently cross-link DNA to any associated proteins provides a means to isolate and identify proteins bound to the DNA at the site of the EdU-biotin label.
Replication Site where DNA synthesis ..
Bambara RA, Murante RS and Henricksen LA (1997) Enzymes and reactions at the eukaryotic DNA replication fork. Journal of Biological Chemistry 272: 4647–4650.