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Inositol phosphates

From duckweed to DNA…

Nature Reviews Molecular Cell Biology volume 1, page 6 (2000)Cite this article

The story revolves around the catalytic subunit of the DNA-dependent protein kinase(DNA-PKcs), which helps to repair double-stranded DNA breaks using the non-homologous end-joining (NHEJ) pathway. The DNA-PKcs is first targeted to each of the severed ends by a complex of the Ku70 and Ku80 proteins (see figure). DNA-PKcs then recruits a complex of DNA ligase IV and XRCC4, resulting in rejoining of the broken ends.

In vitro assays have shown that although all of these players — Ku70, Ku80, DNA-PKcs, XRCC4 and DNA ligase IV — are needed for NHEJ, they are not sufficient. So something is missing, and to discover what, Hanakahi et al. used an in vitro complementation assay. They took cell-free extracts that could promote NHEJ, fractionated them on a phosphocellulose column, then added back various combinations of the fractions to restore the NHEJ activity. In so doing, the authors identified a fraction containing the elusive factor, which they named stimulatory factor A (SFA).

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References

ORIGINAL RESEARCH PAPER

  1. Hanakahi, L. A., Bartlet-Jones, M., Chappell, C., Pappin, D. & West, S. C. Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair. Cell 102, 721–729 (2000)

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REVIEWS

  1. Smith, G. C. M. & Jackson, S. P. The DNA-dependent protein kinase. Genes Dev. 13, 916–934 (1999)

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  2. Shears, S. B. Transcriptional regulation: a new dominion for inositol phosphate signaling? . Bioessays 22, 786–789 (2000)

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  3. Lewis, L. K. & Resnick, M. A. Tying up loose ends: nonhomologous end-joining in saccharomyces cerevisiae. Mutat. Res. 451, 71–89 ( 2000)

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Authors
  1. Alison Mitchell
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Mitchell, A. From duckweed to DNA…. Nat Rev Mol Cell Biol 1, 6 (2000). https://doi.org/10.1038/35036017

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