The annealing helicase SMARCAL1 maintains genome integrity at stalled replication forks

Genes Dev. 2009 Oct 15;23(20):2405-14. doi: 10.1101/gad.1839909. Epub 2009 Sep 30.

Abstract

Mutations in SMARCAL1 (HARP) cause Schimke immunoosseous dysplasia (SIOD). The mechanistic basis for this disease is unknown. Using functional genomic screens, we identified SMARCAL1 as a genome maintenance protein. Silencing and overexpression of SMARCAL1 leads to activation of the DNA damage response during S phase in the absence of any genotoxic agent. SMARCAL1 contains a Replication protein A (RPA)-binding motif similar to that found in the replication stress response protein TIPIN (Timeless-Interacting Protein), which is both necessary and sufficient to target SMARCAL1 to stalled replication forks. RPA binding is critical for the cellular function of SMARCAL1; however, it is not necessary for the annealing helicase activity of SMARCAL1 in vitro. An SIOD-associated SMARCAL1 mutant fails to prevent replication-associated DNA damage from accumulating in cells in which endogenous SMARCAL1 is silenced. Ataxia-telangiectasia mutated (ATM), ATM and Rad3-related (ATR), and DNA-dependent protein kinase (DNA-PK) phosphorylate SMARCAL1 in response to replication stress. Loss of SMARCAL1 activity causes increased RPA loading onto chromatin and persistent RPA phosphorylation after a transient exposure to replication stress. Furthermore, SMARCAL1-deficient cells are hypersensitive to replication stress agents. Thus, SMARCAL1 is a replication stress response protein, and the pleiotropic phenotypes of SIOD are at least partly due to defects in genome maintenance during DNA replication.

Publication types

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

MeSH terms

  • Cell Line
  • Chromatin
  • DNA Helicases / metabolism*
  • DNA Replication*
  • Gene Expression Regulation
  • Genomic Instability*
  • HeLa Cells
  • Humans
  • Mutation
  • Osteochondrodysplasias / genetics
  • Osteochondrodysplasias / physiopathology
  • Phosphorylation
  • Protein Binding
  • Replication Protein A / metabolism

Substances

  • Chromatin
  • Replication Protein A
  • SMARCAL1 protein, human
  • DNA Helicases