Oxidative damage diminishes mitochondrial DNA polymerase replication fidelity.

Abstract:

:Mitochondrial DNA (mtDNA) resides in a high ROS environment and suffers more mutations than its nuclear counterpart. Increasing evidence suggests that mtDNA mutations are not the results of direct oxidative damage, rather are caused, at least in part, by DNA replication errors. To understand how the mtDNA replicase, Pol γ, can give rise to elevated mutations, we studied the effect of oxidation of Pol γ on replication errors. Pol γ is a high fidelity polymerase with polymerase (pol) and proofreading exonuclease (exo) activities. We show that Pol γ exo domain is far more sensitive to oxidation than pol; under oxidative conditions, exonuclease activity therefore declines more rapidly than polymerase. The oxidized Pol γ becomes editing-deficient, displaying a 20-fold elevated mutations than the unoxidized enzyme. Mass spectrometry analysis reveals that Pol γ exo domain is a hotspot for oxidation. The oxidized exo residues increase the net negative charge around the active site that should reduce the affinity to mismatched primer/template DNA. Our results suggest that the oxidative stress induced high mutation frequency on mtDNA can be indirectly caused by oxidation of the mitochondrial replicase.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Anderson AP,Luo X,Russell W,Yin YW

doi

10.1093/nar/gkz1018

subject

Has Abstract

pub_date

2020-01-24 00:00:00

pages

817-829

issue

2

eissn

0305-1048

issn

1362-4962

pii

5651319

journal_volume

48

pub_type

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