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Barrier-to-autointegration factor proteome reveals chromatin-regulatory partners.

Nuclear lamin filaments and associated proteins form a nucleoskeletal ("lamina") network required for transcription, replication, chromatin organization and epigenetic regulation in metazoans. Lamina defects cause human disease ("laminopathies") and are linked to aging. Barrier-to-autointegration factor (BAF) is a mobile and essential component of the nuclear lamina that binds directly to histones, lamins and LEM-domain proteins, including the inner nuclear membrane protein emerin, and has roles in chromatin structure, mitosis and gene regulation. To understand BAF's mechanisms of action, BAF associated proteins were affinity-purified from HeLa cell nuclear lysates using BAF-conjugated beads, and identified by tandem mass spectrometry or independently identified and quantified using the iTRAQ method. We recovered A- and B-type lamins and core histones, all known to bind BAF directly, plus four human transcription factors (Requiem, NonO, p15, LEDGF), disease-linked proteins (e.g., Huntingtin, Treacle) and several proteins and enzymes that regulate chromatin. Association with endogenous BAF was independently validated by co-immunoprecipitation from HeLa cells for seven candidates including Requiem, poly(ADP-ribose) polymerase 1 (PARP1), retinoblastoma binding protein 4 (RBBP4), damage-specific DNA binding protein 1 (DDB1) and DDB2. Interestingly, endogenous BAF and emerin each associated with DDB2 and CUL4A in a UV- and time-dependent manner, suggesting BAF and emerin have dynamic roles in genome integrity and might help couple DNA damage responses to the nuclear lamina network. We conclude this proteome is a rich source of candidate partners for BAF and potentially also A- and B-type lamins, which may reveal how chromatin regulation and genome integrity are linked to nuclear structure.

Pubmed ID: 19759913

Authors

  • Montes de Oca R
  • Shoemaker CJ
  • Gucek M
  • Cole RN
  • Wilson KL

Journal

PloS one

Publication Data

September 17, 2009

Associated Grants

  • Agency: NIGMS NIH HHS, Id: R01 GM48646

Mesh Terms

  • Amino Acid Sequence
  • Chromatin
  • Cullin Proteins
  • DNA-Binding Proteins
  • HeLa Cells
  • Histones
  • Humans
  • Membrane Proteins
  • Molecular Sequence Data
  • Nuclear Lamina
  • Nuclear Proteins
  • Poly(ADP-ribose) Polymerases
  • Protein Structure, Tertiary
  • Proteome
  • Retinoblastoma-Binding Protein 4
  • Sequence Homology, Amino Acid