Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction

The bacterial single-stranded DNA (ssDNA) binding protein SSB is a strictly conserved and essential protein involved in diverse functions of DNA metabolism, including replication and repair. SSB comprises a well-characterized tetrameric core of N-terminal oligonucleotide binding OB folds that bind s...

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Main Authors: Green, Matthew, Hatter, Louise, Brookes, Emre, Soultanas, Panos, Scott, David J.
Format: Article
Language:English
Published: Elsevier 2016
Subjects:
Online Access:https://eprints.nottingham.ac.uk/34212/
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author Green, Matthew
Hatter, Louise
Brookes, Emre
Soultanas, Panos
Scott, David J.
author_facet Green, Matthew
Hatter, Louise
Brookes, Emre
Soultanas, Panos
Scott, David J.
author_sort Green, Matthew
building Nottingham Research Data Repository
collection Online Access
description The bacterial single-stranded DNA (ssDNA) binding protein SSB is a strictly conserved and essential protein involved in diverse functions of DNA metabolism, including replication and repair. SSB comprises a well-characterized tetrameric core of N-terminal oligonucleotide binding OB folds that bind ssDNA and four intrinsically disordered C-terminal domains of unknown structure that interact with partner proteins. The generally accepted, albeit speculative, mechanistic model in the field postulates that binding of ssDNA to the OB core induces the flexible, undefined C-terminal arms to expand outwards encouraging functional interactions with partner proteins. In this structural study, we show that the opposite is true. Combined small-angle scattering with X-rays and neutrons coupled to coarse-grained modeling reveal that the intrinsically disordered C-terminal arms are relatively collapsed around the tetrameric OB core and collapse further upon ssDNA binding. This implies a mechanism of action, in which the disordered C-terminal domain collapse traps the ssDNA and pulls functional partners onto the ssDNA.
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spelling nottingham-342122020-05-08T11:45:27Z https://eprints.nottingham.ac.uk/34212/ Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction Green, Matthew Hatter, Louise Brookes, Emre Soultanas, Panos Scott, David J. The bacterial single-stranded DNA (ssDNA) binding protein SSB is a strictly conserved and essential protein involved in diverse functions of DNA metabolism, including replication and repair. SSB comprises a well-characterized tetrameric core of N-terminal oligonucleotide binding OB folds that bind ssDNA and four intrinsically disordered C-terminal domains of unknown structure that interact with partner proteins. The generally accepted, albeit speculative, mechanistic model in the field postulates that binding of ssDNA to the OB core induces the flexible, undefined C-terminal arms to expand outwards encouraging functional interactions with partner proteins. In this structural study, we show that the opposite is true. Combined small-angle scattering with X-rays and neutrons coupled to coarse-grained modeling reveal that the intrinsically disordered C-terminal arms are relatively collapsed around the tetrameric OB core and collapse further upon ssDNA binding. This implies a mechanism of action, in which the disordered C-terminal domain collapse traps the ssDNA and pulls functional partners onto the ssDNA. Elsevier 2016-01-29 Article PeerReviewed application/pdf en cc_by_nc_nd https://eprints.nottingham.ac.uk/34212/1/JMB-D-15-00738R2.pdf Green, Matthew, Hatter, Louise, Brookes, Emre, Soultanas, Panos and Scott, David J. (2016) Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction. Journal of Molecular Biology, 428 (2). pp. 357-364. ISSN 1089-8638 intrinsic disorder; SASSIE; SANS; SAXS; Bacillus subtilis http://www.sciencedirect.com/science/article/pii/S0022283615006981 doi:10.1016/j.jmb.2015.12.007 doi:10.1016/j.jmb.2015.12.007
spellingShingle intrinsic disorder; SASSIE; SANS; SAXS; Bacillus subtilis
Green, Matthew
Hatter, Louise
Brookes, Emre
Soultanas, Panos
Scott, David J.
Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title_full Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title_fullStr Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title_full_unstemmed Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title_short Defining the intrinsically disordered C-terminal domain of SSB reveals DNA-mediated compaction
title_sort defining the intrinsically disordered c-terminal domain of ssb reveals dna-mediated compaction
topic intrinsic disorder; SASSIE; SANS; SAXS; Bacillus subtilis
url https://eprints.nottingham.ac.uk/34212/
https://eprints.nottingham.ac.uk/34212/
https://eprints.nottingham.ac.uk/34212/