Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity
Mitochondrial DNA (mtDNA) damaged by reactive oxygen species (ROS) triggers so far poorly understood processes of mtDNA maintenance that are coordinated by a complex interplay among DNA repair, DNA degradation, and DNA replication. This study was designed to identify the proteins involved in mtDNA m...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-05-01
|
| Series: | Genes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4425/13/5/879 |
| _version_ | 1797499573394997248 |
|---|---|
| author | Heike Katrin Schreier Rahel Stefanie Wiehe Miria Ricchetti Lisa Wiesmüller |
| author_facet | Heike Katrin Schreier Rahel Stefanie Wiehe Miria Ricchetti Lisa Wiesmüller |
| author_sort | Heike Katrin Schreier |
| collection | DOAJ |
| description | Mitochondrial DNA (mtDNA) damaged by reactive oxygen species (ROS) triggers so far poorly understood processes of mtDNA maintenance that are coordinated by a complex interplay among DNA repair, DNA degradation, and DNA replication. This study was designed to identify the proteins involved in mtDNA maintenance by applying a special long-range PCR, reflecting mtDNA integrity in the minor arc. A siRNA screening of literature-based candidates was performed under conditions of enforced oxidative phosphorylation revealing the functional group of polymerases and therein polymerase ζ (POLZ) as top hits. Thus, POLZ knockdown caused mtDNA accumulation, which required the activity of the base excision repair (BER) nuclease APE1, and was followed by compensatory mtDNA replication determined by the single-cell mitochondrial in situ hybridization protocol (mTRIP). Quenching reactive oxygen species (ROS) in mitochondria unveiled an additional, ROS-independent involvement of POLZ in the formation of a typical deletion in the minor arc region. Together with data demonstrating the localization of POLZ in mitochondria, we suggest that POLZ plays a significant role in mtDNA turnover, particularly under conditions of oxidative stress. |
| first_indexed | 2024-03-10T03:49:19Z |
| format | Article |
| id | doaj.art-735a9c92b58a4b93ba25cc877bd126d0 |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T03:49:19Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-735a9c92b58a4b93ba25cc877bd126d02023-11-23T11:10:58ZengMDPI AGGenes2073-44252022-05-0113587910.3390/genes13050879Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 ActivityHeike Katrin Schreier0Rahel Stefanie Wiehe1Miria Ricchetti2Lisa Wiesmüller3Department of Obstetrics and Gynecology, Ulm University, 89075 Ulm, GermanyDepartment of Obstetrics and Gynecology, Ulm University, 89075 Ulm, GermanyDepartment of Developmental and Stem Cell Biology, Institute Pasteur, CEDEX 15, 75724 Paris, FranceDepartment of Obstetrics and Gynecology, Ulm University, 89075 Ulm, GermanyMitochondrial DNA (mtDNA) damaged by reactive oxygen species (ROS) triggers so far poorly understood processes of mtDNA maintenance that are coordinated by a complex interplay among DNA repair, DNA degradation, and DNA replication. This study was designed to identify the proteins involved in mtDNA maintenance by applying a special long-range PCR, reflecting mtDNA integrity in the minor arc. A siRNA screening of literature-based candidates was performed under conditions of enforced oxidative phosphorylation revealing the functional group of polymerases and therein polymerase ζ (POLZ) as top hits. Thus, POLZ knockdown caused mtDNA accumulation, which required the activity of the base excision repair (BER) nuclease APE1, and was followed by compensatory mtDNA replication determined by the single-cell mitochondrial in situ hybridization protocol (mTRIP). Quenching reactive oxygen species (ROS) in mitochondria unveiled an additional, ROS-independent involvement of POLZ in the formation of a typical deletion in the minor arc region. Together with data demonstrating the localization of POLZ in mitochondria, we suggest that POLZ plays a significant role in mtDNA turnover, particularly under conditions of oxidative stress.https://www.mdpi.com/2073-4425/13/5/879polymerase ζmitochondrial DNA degradationoxidative damagebase excision repair |
| spellingShingle | Heike Katrin Schreier Rahel Stefanie Wiehe Miria Ricchetti Lisa Wiesmüller Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity Genes polymerase ζ mitochondrial DNA degradation oxidative damage base excision repair |
| title | Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity |
| title_full | Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity |
| title_fullStr | Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity |
| title_full_unstemmed | Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity |
| title_short | Polymerase ζ Is Involved in Mitochondrial DNA Maintenance Processes in Concert with APE1 Activity |
| title_sort | polymerase ζ is involved in mitochondrial dna maintenance processes in concert with ape1 activity |
| topic | polymerase ζ mitochondrial DNA degradation oxidative damage base excision repair |
| url | https://www.mdpi.com/2073-4425/13/5/879 |
| work_keys_str_mv | AT heikekatrinschreier polymerasezisinvolvedinmitochondrialdnamaintenanceprocessesinconcertwithape1activity AT rahelstefaniewiehe polymerasezisinvolvedinmitochondrialdnamaintenanceprocessesinconcertwithape1activity AT miriaricchetti polymerasezisinvolvedinmitochondrialdnamaintenanceprocessesinconcertwithape1activity AT lisawiesmuller polymerasezisinvolvedinmitochondrialdnamaintenanceprocessesinconcertwithape1activity |