Translesion DNA synthesis (TLS) involves PCNA mono-ubiquitination and TLS DNA polymerases (pols). Recent evidence has shown that the mono-ubiquitination is induced not only by DNA damage but also by other factors that induce stalling of the DNA replication fork. We studied the effect of spontaneous DNA replication errors on PCNA mono-ubiquitination and TLS induction. In the pol1L868F strain, which expressed an error-prone pol alpha, PCNA was spontaneously mono-ubiquitinated. Pol alpha L868F had a rate-limiting step at the extension from mismatched primer termini. Electron microscopic observation showed the accumulation of a single-stranded region at the DNA replication fork in yeast cells. For pol alpha errors, pol zeta participated in a generation of +1 frameshifts. Furthermore, in the pol1L868F strain, UV-induced mutations were lower than in the wild-type and a pol delta mutant strain (pol3-5DV), and deletion of the RAD30 gene (pol eta) suppressed this defect. These data suggest that nucleotide misincorporation by pol alpha induces exposure of single-stranded DNA, PCNA mono-ubiquitination and activates TLS pols.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3892766 | PMC |
| http://dx.doi.org/10.1093/jb/mvp043 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Rad5 and Ubc4 directly ubiquitinate PCNA at Lys164 in vitro.
J Biol Chem
January 2025
Department of Biochemistry and Molecular Biology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The province and ministry co-sponsored collaborative innovation center for medical epigenetics, Tianjin Key Laboratory of Medical Epigenetics, Tianjin Medical University, 300070 Tianjin, P. R. CHINA. Electronic address:
Ubiquitination of the proliferating cell nuclear antigen (PCNA) by the budding yeast protein Rad5 have important functions in replication stress responses. Rad5 together with the Ubc13-Mms2 complex attaches Lys63-linked ubiquitin chain to a highly conserved Lys164 residue in PCNA. The reaction requires prior PCNA mono-ubiquitination by the Rad6-Rad18 complex and signals for error-free DNA damage tolerance responses.
View Article and Find Full Text PDFDNA polymerase η is regulated by mutually exclusive mono-ubiquitination and mono-NEDDylation.
bioRxiv
October 2024
Laboratory of Genomic Integrity, National Institute of Child Health and Human Development, National Institutes of Health, 9800 Medical Center Drive, Bethesda, MD 20892-3371, USA.
DNA polymerase eta (Pol η) is a Y-family translesion polymerase responsible for synthesizing new DNA across UV-damaged templates. It is recruited to replication forks following mono-ubiquitination of the PCNA DNA clamp. This interaction is mediated by PCNA-interacting protein (PIP) motifs within Pol η, as well as by its C-terminal ubiquitin-binding zinc finger (UBZ) domain.
View Article and Find Full Text PDFStructural basis for RAD18 regulation by MAGEA4 and its implications for RING ubiquitin ligase binding by MAGE family proteins.
EMBO J
April 2024
European Molecular Biology Laboratory, 71 Avenue des Martyrs, 38042, Grenoble, France.
Article Synopsis
- MAGEA4 is a cancer-testis antigen that is normally found in the testes but is also overexpressed in various cancers, where it plays a role in tumor evolution by activating DNA synthesis processes.
- The study utilized NMR and AlphaFold2 to clarify the complex interaction between MAGEA4 and RAD18, showing that MAGEA4 partially displaces RAD6 from RAD18, affecting its autoubiquitination and function.
- Further findings indicate that the interaction pattern observed in the MAGEA4/RAD18 complex may extend to other Type-I MAGE proteins, like MAGE-C2, offering new insights into the regulation of DNA repair processes through PCNA mono-ubiquitination.
FANCD2-dependent mitotic DNA synthesis relies on PCNA K164 ubiquitination.
Cell Rep
December 2023
Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA; Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA 22903, USA. Electronic address:
Article Synopsis
- * Mutations preventing K164 ubiquitination disrupt normal DNA replication and lead to increased replication stress, especially when paired with the DNA polymerase inhibitor aphidicolin.
- * The study highlights that impaired FANCD2 activity in response to K164 mutations reduces its ability to associate with chromatin, thereby hindering the mitotic DNA synthesis process, which is essential to prevent under-replicated DNA.
SUMOylation regulates low-temperature survival and oxidative DNA damage tolerance in Botrytis cinerea.
New Phytol
April 2023
State Key Laboratory of Rice Biology, Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Biotechnology, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, China.
SUMOylation as one of the protein post-translational modifications plays crucial roles in multiple biological processes of eukaryotic organisms. Botrytis cinerea is a devastating fungal pathogen and capable of infecting plant hosts at low temperature. However, the molecular mechanisms of low-temperature adaptation are largely unknown in fungi.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!