Canonical single-stranded DNA-binding proteins (SSBs) from the oligosaccharide/oligonucleotide-binding (OB) domain family are present in all known organisms and are critical for DNA replication, recombination and repair. The SSB from the hyperthermophilic crenarchaeote Sulfolobus solfataricus (SsoSSB) has a ‘simple’ domain organization consisting of a single DNA-binding OB fold coupled to a flexible C-terminal tail, in contrast with other SSBs in this family that incorporate up to four OB domains. Despite the large differences in the domain organization within the SSB family, the structure of the OB domain is remarkably similar all cellular life forms. However, there are significant differences in the molecular mechanism of ssDNA binding. We have determined the structure of the SsoSSB OB domain bound to ssDNA by NMR spectroscopy. We reveal that ssDNA recognition is modulated by base-stacking of three key aromatic residues, in contrast with the OB domains of human RPA and the recently discovered human homologue of SsoSSB, hSSB1. We also demonstrate that SsoSSB binds ssDNA with a footprint of five bases and with a defined binding polarity. These data elucidate the structural basis of DNA binding and shed light on the molecular mechanism by which these ‘simple’ SSBs interact with ssDNA.
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Research Article|
January 06 2015
The structural basis of DNA binding by the single-stranded DNA-binding protein from Sulfolobus solfataricus
Roland Gamsjaeger;
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
†School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia
1Correspondence may be addressed to either of these authors (email r.gamsjaeger@uws.edu.au or l.cubeddu@uws.edu.au).
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Ruvini Kariawasam;
Ruvini Kariawasam
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
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Adrian X. Gimenez;
Adrian X. Gimenez
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
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Christine Touma;
Christine Touma
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
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Elysse McIlwain;
Elysse McIlwain
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
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Ray E. Bernardo;
Ray E. Bernardo
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
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Nicholas E. Shepherd;
Nicholas E. Shepherd
†School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia
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Sandro F. Ataide;
Sandro F. Ataide
†School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia
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Qihan Dong;
Qihan Dong
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
‡Central Clinical School and Bosch Institute, The University of Sydney and Department of Endocrinology, Royal Prince Alfred Hospital, Sydney, Australia
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Derek J. Richard;
Derek J. Richard
§Institute of Health and Biomedical Innovation, Translational Research Institute, Queensland University of Technology, Woolloongabba 4102, Australia
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Malcolm F. White;
Malcolm F. White
║Biomolecular Sciences Research Complex, University of St Andrews, St Andrews, KY16 9ST, U.K.
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Liza Cubeddu
*School of Science and Health, University of Western Sydney, Penrith, NSW 2751, Australia
†School of Molecular Bioscience, University of Sydney, Sydney, NSW 2006, Australia
1Correspondence may be addressed to either of these authors (email r.gamsjaeger@uws.edu.au or l.cubeddu@uws.edu.au).
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Publisher: Portland Press Ltd
Received:
September 09 2014
Revision Received:
October 31 2014
Accepted:
November 04 2014
Accepted Manuscript online:
November 04 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 465 (2): 337–346.
Article history
Received:
September 09 2014
Revision Received:
October 31 2014
Accepted:
November 04 2014
Accepted Manuscript online:
November 04 2014
Citation
Roland Gamsjaeger, Ruvini Kariawasam, Adrian X. Gimenez, Christine Touma, Elysse McIlwain, Ray E. Bernardo, Nicholas E. Shepherd, Sandro F. Ataide, Qihan Dong, Derek J. Richard, Malcolm F. White, Liza Cubeddu; The structural basis of DNA binding by the single-stranded DNA-binding protein from Sulfolobus solfataricus. Biochem J 15 January 2015; 465 (2): 337–346. doi: https://doi.org/10.1042/BJ20141140
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