Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA

The ParB protein, KorB, from the RK2 plasmid is required for DNA partitioning and transcriptional repression. It acts co-operatively with other proteins, including the repressor KorA. Like many multifunctional proteins, KorB contains regions of intrinsically disordered structure, existing in a large...

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Main Authors: Hyde, Eva I., Callow, Phillip, Rajasekar, Karthik V., timmins, Peter, Patel, Trushar Patel, Siligardi, Giuliano, Hussain, Rohanah, White, Scott A., Thomas, Christopher M., Scott, David J.
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Published: Portland Press 2017
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Online Access:https://eprints.nottingham.ac.uk/45548/
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author Hyde, Eva I.
Callow, Phillip
Rajasekar, Karthik V.
timmins, Peter
Patel, Trushar Patel
Siligardi, Giuliano
Hussain, Rohanah
White, Scott A.
Thomas, Christopher M.
Scott, David J.
author_facet Hyde, Eva I.
Callow, Phillip
Rajasekar, Karthik V.
timmins, Peter
Patel, Trushar Patel
Siligardi, Giuliano
Hussain, Rohanah
White, Scott A.
Thomas, Christopher M.
Scott, David J.
author_sort Hyde, Eva I.
building Nottingham Research Data Repository
collection Online Access
description The ParB protein, KorB, from the RK2 plasmid is required for DNA partitioning and transcriptional repression. It acts co-operatively with other proteins, including the repressor KorA. Like many multifunctional proteins, KorB contains regions of intrinsically disordered structure, existing in a large ensemble of interconverting conformations. Using NMR spectroscopy, circular dichroism and small-angle neutron scattering, we studied KorB selectively within its binary complexes with KorA and DNA, and within the ternary KorA/KorB/DNA complex. The bound KorB protein remains disordered with a mobile C-terminal domain and no changes in the secondary structure, but increases in the radius of gyration on complex formation. Comparison of wild-type KorB with an N-terminal deletion mutant allows a model of the ensemble average distances betweenthe domains when bound to DNA. We propose that the positive co-operativity between KorB, KorA and DNA results from conformational restriction of KorB on binding each partner, while maintaining disorder.
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publishDate 2017
publisher Portland Press
recordtype eprints
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spelling nottingham-455482020-05-04T19:04:04Z https://eprints.nottingham.ac.uk/45548/ Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA Hyde, Eva I. Callow, Phillip Rajasekar, Karthik V. timmins, Peter Patel, Trushar Patel Siligardi, Giuliano Hussain, Rohanah White, Scott A. Thomas, Christopher M. Scott, David J. The ParB protein, KorB, from the RK2 plasmid is required for DNA partitioning and transcriptional repression. It acts co-operatively with other proteins, including the repressor KorA. Like many multifunctional proteins, KorB contains regions of intrinsically disordered structure, existing in a large ensemble of interconverting conformations. Using NMR spectroscopy, circular dichroism and small-angle neutron scattering, we studied KorB selectively within its binary complexes with KorA and DNA, and within the ternary KorA/KorB/DNA complex. The bound KorB protein remains disordered with a mobile C-terminal domain and no changes in the secondary structure, but increases in the radius of gyration on complex formation. Comparison of wild-type KorB with an N-terminal deletion mutant allows a model of the ensemble average distances betweenthe domains when bound to DNA. We propose that the positive co-operativity between KorB, KorA and DNA results from conformational restriction of KorB on binding each partner, while maintaining disorder. Portland Press 2017-08-31 Article PeerReviewed Hyde, Eva I., Callow, Phillip, Rajasekar, Karthik V., timmins, Peter, Patel, Trushar Patel, Siligardi, Giuliano, Hussain, Rohanah, White, Scott A., Thomas, Christopher M. and Scott, David J. (2017) Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA. Biochemical Journal, 474 (18). pp. 3121-3135. ISSN 0264-6021 circular dichroism; intrinsically disordered regions; small-angle neutron scattering experiments; small-angle X-ray scattering; http://www.biochemj.org/content/474/18/3121 doi:10.1042/BCJ20170281 doi:10.1042/BCJ20170281
spellingShingle circular dichroism; intrinsically disordered regions; small-angle neutron scattering experiments; small-angle X-ray scattering;
Hyde, Eva I.
Callow, Phillip
Rajasekar, Karthik V.
timmins, Peter
Patel, Trushar Patel
Siligardi, Giuliano
Hussain, Rohanah
White, Scott A.
Thomas, Christopher M.
Scott, David J.
Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title_full Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title_fullStr Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title_full_unstemmed Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title_short Intrinsic disorder in the partitioning protein KorB persists after co-operative complex formation with operator DNA and KorA
title_sort intrinsic disorder in the partitioning protein korb persists after co-operative complex formation with operator dna and kora
topic circular dichroism; intrinsically disordered regions; small-angle neutron scattering experiments; small-angle X-ray scattering;
url https://eprints.nottingham.ac.uk/45548/
https://eprints.nottingham.ac.uk/45548/
https://eprints.nottingham.ac.uk/45548/