Charting the landscape of tandem BRCT domain-mediated protein interactions

Sci Signal. 2012 Sep 18;5(242):rs6. doi: 10.1126/scisignal.2002255.

Abstract

Eukaryotic cells have evolved an intricate system to resolve DNA damage to prevent its transmission to daughter cells. This system, collectively known as the DNA damage response (DDR) network, includes many proteins that detect DNA damage, promote repair, and coordinate progression through the cell cycle. Because defects in this network can lead to cancer, this network constitutes a barrier against tumorigenesis. The modular BRCA1 carboxyl-terminal (BRCT) domain is frequently present in proteins involved in the DDR, can exist either as an individual domain or as tandem domains (tBRCT), and can bind phosphorylated peptides. We performed a systematic analysis of protein-protein interactions involving tBRCT in the DDR by combining literature curation, yeast two-hybrid screens, and tandem affinity purification coupled to mass spectrometry. We identified 23 proteins containing conserved BRCT domains and generated a human protein-protein interaction network for seven proteins with tBRCT. This study also revealed previously unknown components in DNA damage signaling, such as COMMD1 and the target of rapamycin complex mTORC2. Additionally, integration of tBRCT domain interactions with DDR phosphoprotein studies and analysis of kinase-substrate interactions revealed signaling subnetworks that may aid in understanding the involvement of tBRCT in disease and DNA repair.

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

  • Adaptor Proteins, Signal Transducing* / genetics
  • Adaptor Proteins, Signal Transducing* / metabolism
  • Animals
  • BRCA1 Protein* / genetics
  • BRCA1 Protein* / metabolism
  • DNA Damage*
  • DNA Repair / physiology*
  • Humans
  • Mechanistic Target of Rapamycin Complex 2
  • Multiprotein Complexes* / genetics
  • Multiprotein Complexes* / metabolism
  • Peptides / genetics
  • Peptides / metabolism
  • Phosphorylation / genetics
  • Protein Structure, Tertiary
  • Sequence Analysis, Protein*
  • TOR Serine-Threonine Kinases* / genetics
  • TOR Serine-Threonine Kinases* / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • BRCA1 Protein
  • BRCA1 protein, human
  • COMMD1 protein, human
  • Multiprotein Complexes
  • Peptides
  • Mechanistic Target of Rapamycin Complex 2
  • TOR Serine-Threonine Kinases