The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction

Summary: The partner and localizer of BRCA2 (PALB2) is a scaffold protein linking BRCA1 with BRCA2 and RAD51 during homologous recombination (HR). PALB2 interaction with DNA strongly enhances HR in cells, while the PALB2 DNA-binding domain (PALB2-DBD) supports DNA strand exchange in vitro. We determ...

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Main Authors: Yevhenii Kyriukha, Maxwell B. Watkins, Jennifer M. Redington, Nithya Chintalapati, Abhishek Ganti, Reza Dastvan, Vladimir N. Uversky, Jesse B. Hopkins, Nicola Pozzi, Sergey Korolev
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:iScience
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589004224024842
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author Yevhenii Kyriukha
Maxwell B. Watkins
Jennifer M. Redington
Nithya Chintalapati
Abhishek Ganti
Reza Dastvan
Vladimir N. Uversky
Jesse B. Hopkins
Nicola Pozzi
Sergey Korolev
author_facet Yevhenii Kyriukha
Maxwell B. Watkins
Jennifer M. Redington
Nithya Chintalapati
Abhishek Ganti
Reza Dastvan
Vladimir N. Uversky
Jesse B. Hopkins
Nicola Pozzi
Sergey Korolev
author_sort Yevhenii Kyriukha
collection DOAJ
description Summary: The partner and localizer of BRCA2 (PALB2) is a scaffold protein linking BRCA1 with BRCA2 and RAD51 during homologous recombination (HR). PALB2 interaction with DNA strongly enhances HR in cells, while the PALB2 DNA-binding domain (PALB2-DBD) supports DNA strand exchange in vitro. We determined that PALB2-DBD is intrinsically disordered beyond a single N-terminal α-helix. Coiled-coil mediated dimerization is stabilized by interaction between intrinsically disordered regions (IDRs) leading to a 2-fold structural compaction. Single-stranded (ss)DNA binding promotes additional structural compaction and protein tetramerization. Using confocal single-molecule FRET, we observed bimodal and oligomerization-dependent compaction of ssDNA bound to PALB2-DBD, suggesting a novel strand exchange mechanism. Bioinformatics analysis and preliminary observations indicate that PALB2 forms protein-nucleic acids condensates. Intrinsically disordered DBDs are prevalent in the human proteome. PALB2-DBD and similar IDRs may use a chaperone-like mechanism to aid formation and resolution of DNA and RNA multichain intermediates during DNA replication, repair and recombination.
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spelling doaj-art-c439c487940a4e6fa6c652461f0b310b2025-08-20T01:57:08ZengElsevieriScience2589-00422024-12-01271211125910.1016/j.isci.2024.111259The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compactionYevhenii Kyriukha0Maxwell B. Watkins1Jennifer M. Redington2Nithya Chintalapati3Abhishek Ganti4Reza Dastvan5Vladimir N. Uversky6Jesse B. Hopkins7Nicola Pozzi8Sergey Korolev9Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USAThe Biophysics Collaborative Access Team (BioCat), Departments of Biology and Physics, Illinois Institute of Technology, Chicago, IL, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USADepartment of Molecular Medicine and USF Health Byrd Alzheimer’s Research Institute, Morsani College of Medicine, University of South Florida, Tampa, FL, USAThe Biophysics Collaborative Access Team (BioCat), Departments of Biology and Physics, Illinois Institute of Technology, Chicago, IL, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USAEdward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, USA; Corresponding authorSummary: The partner and localizer of BRCA2 (PALB2) is a scaffold protein linking BRCA1 with BRCA2 and RAD51 during homologous recombination (HR). PALB2 interaction with DNA strongly enhances HR in cells, while the PALB2 DNA-binding domain (PALB2-DBD) supports DNA strand exchange in vitro. We determined that PALB2-DBD is intrinsically disordered beyond a single N-terminal α-helix. Coiled-coil mediated dimerization is stabilized by interaction between intrinsically disordered regions (IDRs) leading to a 2-fold structural compaction. Single-stranded (ss)DNA binding promotes additional structural compaction and protein tetramerization. Using confocal single-molecule FRET, we observed bimodal and oligomerization-dependent compaction of ssDNA bound to PALB2-DBD, suggesting a novel strand exchange mechanism. Bioinformatics analysis and preliminary observations indicate that PALB2 forms protein-nucleic acids condensates. Intrinsically disordered DBDs are prevalent in the human proteome. PALB2-DBD and similar IDRs may use a chaperone-like mechanism to aid formation and resolution of DNA and RNA multichain intermediates during DNA replication, repair and recombination.http://www.sciencedirect.com/science/article/pii/S2589004224024842BiochemistryMolecular biologyCell biology
spellingShingle Yevhenii Kyriukha
Maxwell B. Watkins
Jennifer M. Redington
Nithya Chintalapati
Abhishek Ganti
Reza Dastvan
Vladimir N. Uversky
Jesse B. Hopkins
Nicola Pozzi
Sergey Korolev
The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
iScience
Biochemistry
Molecular biology
Cell biology
title The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
title_full The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
title_fullStr The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
title_full_unstemmed The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
title_short The strand exchange domain of tumor suppressor PALB2 is intrinsically disordered and promotes oligomerization-dependent DNA compaction
title_sort strand exchange domain of tumor suppressor palb2 is intrinsically disordered and promotes oligomerization dependent dna compaction
topic Biochemistry
Molecular biology
Cell biology
url http://www.sciencedirect.com/science/article/pii/S2589004224024842
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