Role of BRCA2 DNA-binding and C-terminal domain on its mobility and conformation in DNA repair

Paul, M. W.; Sidhu, A.; Liang, Y.; van Rossum-Fikkert, S. E.; Odijk, H.; Zelensky, A. N.; Kanaar, R.; Wyman, C.

doi:10.1101/2021.03.02.433541

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bioRxiv · 10.1101/2021.03.02.433541

Role of BRCA2 DNA-binding and C-terminal domain on its mobility and conformation in DNA repair

Paul, M. W.·Sidhu, A.·Liang, Y.·van Rossum-Fikkert, S. E.·Odijk, H.·Zelensky, A. N.·Kanaar, R.·Wyman, C.

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Abstract

BRCA2 is an essential protein in genome maintenance, homologous recombination and replication fork protection. Its function includes multiple interaction partners and requires timely localization to relevant sites in the nucleus. We investigated the importance of the highly conserved DNA binding domain (DBD) and C-terminal domain (CTD) of BRCA2. We generated BRCA2 variants missing one or both domains in mouse ES cells and defined their contribution in HR function and dynamic localization in the nucleus, by single particle tracking of BRCA2 mobility. Changes in molecular architecture of BRCA2 induced by binding partners of purified BRCA2 was determined by scanning force microscopy. BRCA2 mobility and DNA damage-induced increase in the immobile fraction was largely unaffected by C- terminal deletions. The purified proteins missing CTD and/or DBD were defective in architectural changes correlating with reduced homologous recombination function in cells. These results emphasize BRCA2 activity at sites of damage beyond promoting RAD51 delivery.

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Paul, M. W., Sidhu, A., Liang, Y., van Rossum-Fikkert, S. E., Odijk, H., Zelensky, A. N., Kanaar, R., Wyman, C.. 2021-03-02. Role of BRCA2 DNA-binding and C-terminal domain on its mobility and conformation in DNA repair. https://doi.org/10.1101/2021.03.02.433541

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