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Preparation of the Mgm101 Recombination Protein by MBP-based Tagging Strategy
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Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity.

Heike Katrin Schreier1, Rahel Stefanie Wiehe1, Miria Ricchetti2

  • 1Department of Obstetrics and Gynecology, Ulm University, 89075 Ulm, Germany.

Genes
|May 28, 2022
PubMed
Summary
This summary is machine-generated.

Polymerase ζ (POLZ) is crucial for mitochondrial DNA (mtDNA) maintenance, especially under oxidative stress. Its knockdown leads to mtDNA accumulation, highlighting its role in DNA repair and replication.

Keywords:
base excision repairmitochondrial DNA degradationoxidative damagepolymerase ζ

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Area of Science:

  • Mitochondrial biology
  • Molecular genetics
  • DNA repair mechanisms

Background:

  • Mitochondrial DNA (mtDNA) damage from reactive oxygen species (ROS) initiates complex maintenance processes involving DNA repair, degradation, and replication.
  • The precise molecular players coordinating mtDNA maintenance under oxidative stress remain incompletely understood.

Purpose of the Study:

  • To identify key proteins involved in mitochondrial DNA maintenance.
  • To elucidate the role of polymerase ζ (POLZ) in mtDNA integrity and turnover.

Main Methods:

  • Utilized long-range PCR to assess mtDNA integrity.
  • Performed siRNA screening targeting candidate proteins under enforced oxidative phosphorylation.
  • Employed single-cell mitochondrial in situ hybridization protocol (mTRIP) to analyze mtDNA replication.
  • Investigated the effect of ROS quenching on POLZ function.

Main Results:

  • siRNA screening identified polymerases, specifically polymerase ζ (POLZ), as critical for mtDNA maintenance.
  • POLZ knockdown resulted in mtDNA accumulation, dependent on APE1 activity, and triggered compensatory mtDNA replication.
  • ROS quenching revealed a ROS-independent role for POLZ in forming specific mtDNA deletions.
  • POLZ was localized to mitochondria, confirming its direct involvement in mitochondrial processes.

Conclusions:

  • Polymerase ζ (POLZ) plays a significant role in mitochondrial DNA turnover.
  • POLZ is essential for maintaining mtDNA integrity, particularly under conditions of oxidative stress.
  • The study implicates POLZ in a complex interplay of mtDNA repair, replication, and degradation pathways.