Purification of Proteins on Newly Synthesized DNA Using Ipond
Document Type
Book Chapter
Publication Date
2015
Keywords
DNA Replication, Chromatin, DNA Repair, Ipond, Edu, Click Chemistry, Replication Stress, PCNA, DNA Damage
Digital Object Identifier (DOI)
https://doi.org/10.1007/978-1-4939-1680-1_10
Abstract
The replisome is a large protein machine containing multiple enzymatic activities needed to complete DNA replication. In addition to helicase and polymerases needed for copying the DNA, the replisome also contains proteins like DNA methyltransferases, histone chaperones, and chromatin modifying enzymes to couple DNA replication with chromatin deposition and establishment of the epigenetic code. In addition, since template DNA strands often contain DNA damage or other roadblocks to the replication machinery, replication stress response proteins associate with the replisome to stabilize, repair, and restart stalled replication forks. Hundreds of proteins are needed to accomplish these tasks. Identifying these proteins, monitoring their posttranslational modifications, and understanding how their activities are coordinated is essential to understand how the genome and epigenome are duplicated rapidly, completely, and accurately every cell division cycle. Here we describe an updated iPOND (isolation of proteins on nascent DNA) method to facilitate these analyses.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Purification of Proteins on Newly Synthesized DNA Using Ipond, in R. Hancock (Ed), The Nucleus. Methods in Molecular Biology, v. 1228, Humana Press, p. 123-131
Scholar Commons Citation
Dungrawala, Huzefa and Cortez, David, "Purification of Proteins on Newly Synthesized DNA Using Ipond" (2015). Molecular Biosciences Faculty Publications. 112.
https://digitalcommons.usf.edu/bcm_facpub/112
