Molecular basis for DNA repair synthesis on short gaps by mycobacterial Primase-Polymerase C

Nigel Brissett; Katerina  Zabrady; Przemyslaw  Plocinski; Julie  Bianchi; Małgorzata  Korycka-Machała; Anna  Brzostek; Jarosław  Dziadek; Aidan J.  Doherty

doi:10.1038/s41467-020-18012-8

Molecular basis for DNA repair synthesis on short gaps by mycobacterial Primase-Polymerase C

Nigel Brissett, Katerina Zabrady, Przemyslaw Plocinski, Julie Bianchi, Małgorzata Korycka-Machała, Anna Brzostek, Jarosław Dziadek, Aidan J. Doherty

Research output: Contribution to journal › Article › peer-review

Abstract

Cells utilise specialized polymerases from the Primase-Polymerase (Prim-Pol) superfamily to maintain genome stability. Prim-Pol's function in genome maintenance pathways including replication, repair and damage tolerance. Mycobacteria contain multiple Prim-Pols required for lesion repair, including Prim-PolC that performs short gap repair synthesis during excision repair. To understand the molecular basis of Prim-PolC's gap recognition and synthesis activities, we elucidated crystal structures of pre- and post-catalytic complexes bound to gapped DNA substrates. These intermediates explain its binding preference for short gaps and reveal a distinctive modus operandi called Synthesis-dependent Template Displacement (STD). This mechanism enables Prim-PolC to couple primer extension with template base dislocation, ensuring that the unpaired templating bases in the gap are ushered into the active site in an ordered manner. Insights provided by these structures establishes the molecular basis of Prim-PolC's gap recognition and extension activities, while also illuminating the mechanisms of primer extension utilised by closely related Prim-Pols.

Original language	English
Article number	4196
Journal	Nature Communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-18012-8
Publication status	Published - 21 Aug 2020

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This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation,
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abstract = "Cells utilise specialized polymerases from the Primase-Polymerase (Prim-Pol) superfamily to maintain genome stability. Prim-Pol's function in genome maintenance pathways including replication, repair and damage tolerance. Mycobacteria contain multiple Prim-Pols required for lesion repair, including Prim-PolC that performs short gap repair synthesis during excision repair. To understand the molecular basis of Prim-PolC's gap recognition and synthesis activities, we elucidated crystal structures of pre- and post-catalytic complexes bound to gapped DNA substrates. These intermediates explain its binding preference for short gaps and reveal a distinctive modus operandi called Synthesis-dependent Template Displacement (STD). This mechanism enables Prim-PolC to couple primer extension with template base dislocation, ensuring that the unpaired templating bases in the gap are ushered into the active site in an ordered manner. Insights provided by these structures establishes the molecular basis of Prim-PolC's gap recognition and extension activities, while also illuminating the mechanisms of primer extension utilised by closely related Prim-Pols.",

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Molecular basis for DNA repair synthesis on short gaps by mycobacterial Primase-Polymerase C

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