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Single-stranded DNA-binding protein hSSB1 is critical for genomic stability

Nature volume 453, pages 677–681 (2008)Cite this article

Abstract

Single-strand DNA (ssDNA)-binding proteins (SSBs) are ubiquitous and essential for a wide variety of DNA metabolic processes, including DNA replication, recombination, DNA damage detection and repair1. SSBs have multiple roles in binding and sequestering ssDNA, detecting DNA damage, stimulating nucleases, helicases and strand-exchange proteins, activating transcription and mediating protein–protein interactions. In eukaryotes, the major SSB, replication protein A (RPA), is a heterotrimer1. Here we describe a second human SSB (hSSB1), with a domain organization closer to the archaeal SSB than to RPA. Ataxia telangiectasia mutated (ATM) kinase phosphorylates hSSB1 in response to DNA double-strand breaks (DSBs). This phosphorylation event is required for DNA damage-induced stabilization of hSSB1. Upon induction of DNA damage, hSSB1 accumulates in the nucleus and forms distinct foci independent of cell-cycle phase. These foci co-localize with other known repair proteins. In contrast to RPA, hSSB1 does not localize to replication foci in S-phase cells and hSSB1 deficiency does not influence S-phase progression. Depletion of hSSB1 abrogates the cellular response to DSBs, including activation of ATM and phosphorylation of ATM targets after ionizing radiation. Cells deficient in hSSB1 exhibit increased radiosensitivity, defective checkpoint activation and enhanced genomic instability coupled with a diminished capacity for DNA repair. These findings establish that hSSB1 influences diverse endpoints in the cellular DNA damage response.

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Figure 1: ATM-dependent stabilization and phosphorylation of hSSB1 after ionizing radiation.
Figure 2: hSSB1 localizes to DNA repair foci after ionizing radiation.
Figure 3: hSSB1 inhibition results in defective ionizing-radiation-induced checkpoint activation and defective ATM signalling.
Figure 4: hSSB1 is required for efficient DNA repair.

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Primary accessions

Gene Expression Omnibus

Data deposits

The genes hSSB1 and hSSB2 are deposited in NCBI under accession numbers NM_024068 and NM_001031716, respectively.

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Acknowledgements

This study was supported by grants from the National Health and Medical Research Council of Australia (to K.K.K.), Cancer Research UK (to S.C.W.), the Biotechnology and Biological Sciences Research Council of the UK (to M.F.W), the National Institutes of Health (CA92245 to J.G.; CA10445 and CA123232 to T.K.P.) and the American Cancer Society (RGS-04-173-01-CCG to T.T.P.).

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Authors and Affiliations

  1. Signal Transduction Laboratory, Queensland Institute of Medical Research, Brisbane, Queensland 4029, Australia,

    Derek J. Richard, Emma Bolderson, Kienan Savage, Sergie Tsvetanov & Kum Kum Khanna

  2. Centre for Biomolecular Sciences, University of St Andrews, North Haugh, St Andrews, Fife KY16 9ST, UK,

    Liza Cubeddu, Ross I. M. Wadsworth & Malcolm F. White

  3. School of Molecular and Microbial Biosciences, University of Sydney, Sydney, New South Wales 2006, Australia,

    Liza Cubeddu

  4. Central Clinical Division, School of Medicine, University of Queensland, Queensland 4072, Australia

    Kienan Savage

  5. Department of Radiation Oncology, Washington University School of Medicine, St Louis, Missouri 63108, USA,

    Girdhar G. Sharma, Raj K. Pandita & Tej K. Pandita

  6. Department of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, Texas 78712, USA,

    Matthew L. Nicolette & Tanya T. Paull

  7. London Research Institute, Clare Hall Laboratories, Cancer Research UK, South Mimms, Hertfordshire EN6 3LD, U.K,

    Michael J. McIlwraith & Stephen C. West

  8. Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan

    Shunichi Takeda

  9. Division of Gene Regulation and Expression, Wellcome Biocentre, University of Dundee, Dundee DD1 5EH, U.K.

    Ronald T. Hay

  10. Institute for Cancer Genetics, Columbia University Medical Center, New York, New York 10032, USA,

    Jean Gautier

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Corresponding authors

Correspondence to Malcolm F. White or Kum Kum Khanna.

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Richard, D., Bolderson, E., Cubeddu, L. et al. Single-stranded DNA-binding protein hSSB1 is critical for genomic stability. Nature 453, 677–681 (2008). https://doi.org/10.1038/nature06883

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