A proteomic characterization of factors enriched at nascent DNA molecules

Cell Rep. 2013 Apr 25;3(4):1105-16. doi: 10.1016/j.celrep.2013.03.009. Epub 2013 Mar 28.

Abstract

DNA replication is facilitated by multiple factors that concentrate in the vicinity of replication forks. Here, we developed an approach that combines the isolation of proteins on nascent DNA chains with mass spectrometry (iPOND-MS), allowing a comprehensive proteomic characterization of the human replisome and replisome-associated factors. In addition to known replisome components, we provide a broad list of proteins that reside in the vicinity of the replisome, some of which were not previously associated with replication. For instance, our data support a link between DNA replication and the Williams-Beuren syndrome and identify ZNF24 as a replication factor. In addition, we reveal that SUMOylation is widespread for factors that concentrate near replisomes, which contrasts with lower UQylation levels at these sites. This resource provides a panoramic view of the proteins that concentrate in the surroundings of the replisome, which should facilitate future investigations on DNA replication and genome maintenance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • DNA / metabolism*
  • DNA Replication
  • Gene Regulatory Networks
  • HEK293 Cells
  • Humans
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / metabolism
  • Mass Spectrometry
  • Proteomics*
  • Small Ubiquitin-Related Modifier Proteins / metabolism
  • Ubiquitination
  • Williams Syndrome / genetics
  • Williams Syndrome / metabolism

Substances

  • Kruppel-Like Transcription Factors
  • Small Ubiquitin-Related Modifier Proteins
  • ZNF24 protein, human
  • DNA