A genome-wide camptothecin sensitivity screen identifies a mammalian MMS22L-NFKBIL2 complex required for genomic stability

Mol Cell. 2010 Nov 24;40(4):645-57. doi: 10.1016/j.molcel.2010.10.022. Epub 2010 Nov 4.

Abstract

Replication stress involving collision of replisomes with camptothecin (CPT)-stabilized DNA-Topoisomerase I adducts activates an ATR-dependent pathway to promote repair by homologous recombination. To identify human genes that protect cells from such replication stress, we performed a genome-wide CPT sensitivity screen. Among numerous candidate genes are two previously unstudied proteins: the ankyrin repeat protein NFKBIL2 and C6ORF167 (MMS22L), distantly related to yeast replication stress regulator Mms22p. MMS22L and NFKBIL2 interact with each other and with FACT (facilitator of chromatin transcription) and MCM (minichromosome maintenance) complexes. Cells depleted of NFKBIL2 or MMS22L are sensitive to DNA-damaging agents, load phosphorylated RPA onto chromatin in a CTIP-dependent manner, activate the ATR/ATRIP-CHK1 and double-strand break repair signaling pathways, and are defective in HR. This study identifies MMS22L-NFKBIL2 as components of the replication stress control pathway and provides a resource for discovery of additional components of this pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Camptothecin / pharmacology*
  • DNA Damage
  • DNA Repair / drug effects
  • DNA Replication / drug effects
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed DNA Polymerase / metabolism
  • Drug Resistance, Neoplasm / drug effects
  • Genetic Testing*
  • Genome, Human / genetics*
  • Genomic Instability / drug effects*
  • HeLa Cells
  • Histones / metabolism
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Multienzyme Complexes / metabolism
  • NF-kappa B / deficiency
  • NF-kappa B / metabolism*
  • Nuclear Proteins / metabolism*
  • Phosphorylation / drug effects
  • Protein Binding / drug effects
  • RNA, Small Interfering / metabolism
  • Recombination, Genetic / drug effects
  • Recombination, Genetic / genetics
  • Replication Protein A / metabolism
  • Reproducibility of Results
  • Stress, Physiological / drug effects
  • Tumor Suppressor p53-Binding Protein 1

Substances

  • DNA-Binding Proteins
  • H2AX protein, human
  • Histones
  • Intracellular Signaling Peptides and Proteins
  • MMS22L protein, human
  • Multienzyme Complexes
  • NF-kappa B
  • Nuclear Proteins
  • RNA, Small Interfering
  • Replication Protein A
  • TONSL protein, human
  • TP53BP1 protein, human
  • Tumor Suppressor p53-Binding Protein 1
  • DNA synthesome
  • DNA-Directed DNA Polymerase
  • RPA2 protein, human
  • Camptothecin