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Nucl._Acids_Res.-2014-Keen-nar_gku214.pdf (4.67 MB)

Molecular dissection of the domain architecture and catalytic activities of human PrimPol

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posted on 2023-06-08, 16:16 authored by Benjamin A Keen, Stanislaw K Jozwiakowski, Laura BaileyLaura Bailey, Julie Bianchi, Aidan DohertyAidan Doherty
PrimPol is a primase–polymerase involved in nuclear and mitochondrial DNA replication in eukaryotic cells. Although PrimPol is predicted to possess an archaeo-eukaryotic primase and a UL52-like zinc finger domain, the role of these domains has not been established. Here, we report that the proposed zinc finger domain of human PrimPol binds zinc ions and is essential for maintaining primase activity. Although apparently dispensable for its polymerase activity, the zinc finger also regulates the processivity and fidelity of PrimPol's extension activities. When the zinc finger is disrupted, PrimPol becomes more promutagenic, has an altered translesion synthesis spectrum and is capable of faithfully bypassing cyclobutane pyrimidine dimer photolesions. PrimPol's polymerase domain binds to both single- and double-stranded DNA, whilst the zinc finger domain binds only to single-stranded DNA. We additionally report that although PrimPol's primase activity is required to restore wild-type replication fork rates in irradiated PrimPol-/- cells, polymerase activity is sufficient to maintain regular replisome progression in unperturbed cells. Together, these findings provide the first analysis of the molecular architecture of PrimPol, describing the activities associated with, and interplay between, its functional domains and defining the requirement for its primase and polymerase activities during nuclear DNA replication.

Funding

BB/F01645X/1; BBSRC

BB/H019723/1; BBSRC

G080130; MRC

History

Publication status

  • Published

File Version

  • Published version

Journal

Nucleic Acids Research

ISSN

0305-1048

Publisher

Oxford University Press

Issue

9

Volume

42

Page range

5830-5845

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2014-04-14

First Open Access (FOA) Date

2014-07-01

First Compliant Deposit (FCD) Date

2014-07-01

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