Nascent DNA Proteomics Reveals a Chromatin Remodeler Required for Topoisomerase I Loading at Replication Forks

Cell Rep. 2016 Apr 12;15(2):300-9. doi: 10.1016/j.celrep.2016.03.027. Epub 2016 Mar 31.

Abstract

During transcription and DNA replication, the DNA template is overwound ahead of RNA and DNA polymerases and relaxed by DNA topoisomerases. Inhibitors of topoisomerases are potent anti-cancer agents. Camptothecin traps topoisomerase I on DNA and exerts preferential cytotoxicity toward cancer cells by way of its interference with the progression of replication forks. Starting with an unbiased proteomic analysis, we find that the chromatin remodeling complex BAZ1B-SMARCA5 accumulates near replication forks in camptothecin-exposed cells. We report that BAZ1B associates with topoisomerase I and facilitates its access to replication forks. Single-molecule analyses of replication structures show that BAZ1B contributes to replication interference by camptothecin. A lack of BAZ1B confers increased cellular tolerance of camptothecin. These findings reveal BAZ1B as a key facilitator of topoisomerase I function during DNA replication that affects the response of cancer cells to topoisomerase I inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Camptothecin / pharmacology
  • Chromatin Assembly and Disassembly* / drug effects
  • Chromosomal Proteins, Non-Histone / metabolism
  • DNA / metabolism*
  • DNA Replication* / drug effects
  • DNA Topoisomerases, Type I / metabolism*
  • HeLa Cells
  • Humans
  • Male
  • Proteomics / methods*
  • Topoisomerase I Inhibitors / pharmacology
  • Transcription Factors / metabolism
  • Transcription, Genetic / drug effects

Substances

  • BAZ1B protein, human
  • Chromosomal Proteins, Non-Histone
  • Topoisomerase I Inhibitors
  • Transcription Factors
  • DNA
  • Adenosine Triphosphatases
  • SMARCA5 protein, human
  • DNA Topoisomerases, Type I
  • TOP1 protein, human
  • Camptothecin