DNA-damage tolerance through PCNA ubiquitination and sumoylation

2020 ◽  
Vol 477 (14) ◽  
pp. 2655-2677
Author(s):  
Li Fan ◽  
Tonghui Bi ◽  
Linxiao Wang ◽  
Wei Xiao
Keyword(s):  
Dna Damage ◽  
Posttranslational Modifications ◽  
Damage Tolerance ◽  
Nuclear Antigen ◽  
Replication Forks ◽  
Dna Damage Tolerance ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Damaging Agents ◽  
Template Switch ◽  
Proliferating Cell

DNA-damage tolerance (DDT) is employed by eukaryotic cells to bypass replication-blocking lesions induced by DNA-damaging agents. In budding yeast Saccharomyces cerevisiae, DDT is mediated by RAD6 epistatic group genes and the central event for DDT is sequential ubiquitination of proliferating cell nuclear antigen (PCNA), a DNA clamp required for replication and DNA repair. DDT consists of two parallel pathways: error-prone DDT is mediated by PCNA monoubiquitination, which recruits translesion synthesis DNA polymerases to bypass lesions with decreased fidelity; and error-free DDT is mediated by K63-linked polyubiquitination of PCNA at the same residue of monoubiquitination, which facilitates homologous recombination-mediated template switch. Interestingly, the same PCNA residue is also subjected to sumoylation, which leads to inhibition of unwanted recombination at replication forks. All three types of PCNA posttranslational modifications require dedicated conjugating and ligation enzymes, and these enzymes are highly conserved in eukaryotes, from yeast to human.

scholarly journals Helicase-Like Transcription Factor HLTF and E3 Ubiquitin Ligase SHPRH Confer DNA Damage Tolerance through Direct Interactions with Proliferating Cell Nuclear Antigen (PCNA)

2020 ◽  
Vol 21 (3) ◽  
pp. 693 ◽  
Author(s):  
Mareike Seelinger ◽  
Marit Otterlei
Keyword(s):  
Dna Damage ◽  
Transcription Factor ◽  
Damage Tolerance ◽  
Genome Instability ◽  
Ring Finger ◽  
Nuclear Antigen ◽  
Common Mutation ◽  
Dna Damage Tolerance ◽  
Replicative Stress ◽  
Template Switch

To prevent replication fork collapse and genome instability under replicative stress, DNA damage tolerance (DDT) mechanisms have evolved. The RAD5 homologs, HLTF (helicase-like transcription factor) and SHPRH (SNF2, histone-linker, PHD and RING finger domain-containing helicase), both ubiquitin ligases, are involved in several DDT mechanisms; DNA translesion synthesis (TLS), fork reversal/remodeling and template switch (TS). Here we show that these two human RAD5 homologs contain functional APIM PCNA interacting motifs. Our results show that both the role of HLTF in TLS in HLTF overexpressing cells, and nuclear localization of SHPRH, are dependent on interaction of HLTF and SHPRH with PCNA. Additionally, we detected multiple changes in the mutation spectra when APIM in overexpressed HLTF or SHPRH were mutated compared to overexpressed wild type proteins. In plasmids from cells overexpressing the APIM mutant version of HLTF, we observed a decrease in C to T transitions, the most common mutation caused by UV irradiation, and an increase in mutations on the transcribed strand. These results strongly suggest that direct binding of HLTF and SHPRH to PCNA is vital for their function in DDT.

scholarly journals Functional and Physical Interaction of Yeast Mgs1 with PCNA: Impact on RAD6-Dependent DNA Damage Tolerance

2006 ◽  
Vol 26 (14) ◽  
pp. 5509-5517 ◽  
Author(s):  
Takashi Hishida ◽  
Tomoko Ohya ◽  
Yoshino Kubota ◽  
Yusuke Kamada ◽  
Hideo Shinagawa
Keyword(s):  
Dna Damage ◽  
Posttranslational Modifications ◽  
Cell Cycle Progression ◽  
Damage Tolerance ◽  
Genome Instability ◽  
Dna Helicase ◽  
Nuclear Antigen ◽  
Physical Interaction ◽  
Dna Damage Tolerance ◽  
Exogenous Dna

ABSTRACT Proliferating cell nuclear antigen (PCNA), a sliding clamp required for processive DNA synthesis, provides attachment sites for various other proteins that function in DNA replication, DNA repair, cell cycle progression and chromatin assembly. It has been shown that differential posttranslational modifications of PCNA by ubiquitin or SUMO play a pivotal role in controlling the choice of pathway for rescuing stalled replication forks. Here, we explored the roles of Mgs1 and PCNA in replication fork rescue. We provide evidence that Mgs1 physically associates with PCNA and that Mgs1 helps suppress the RAD6 DNA damage tolerance pathway in the absence of exogenous DNA damage. We also show that PCNA sumoylation inhibits the growth of mgs1 rad18 double mutants, in which PCNA sumoylation and the Srs2 DNA helicase coordinately prevent RAD52-dependent homologous recombination. The proposed roles for Mgs1, Srs2, and modified PCNA during replication arrest highlight the importance of modulating the RAD6 and RAD52 pathways to avoid genome instability.

scholarly journals Control of PCNA deubiquitylation in yeast

2010 ◽  
Vol 38 (1) ◽  
pp. 104-109 ◽  
Author(s):  
Alfonso Gallego-Sánchez ◽  
Francisco Conde ◽  
Pedro San Segundo ◽  
Avelino Bueno
Keyword(s):  
Dna Damage ◽  
Crucial Role ◽  
Proliferating Cell Nuclear Antigen ◽  
Molecular Mechanisms ◽  
Damage Tolerance ◽  
Nuclear Antigen ◽  
Dna Damage Tolerance ◽  
Sliding Clamp ◽  
Evolutionarily Conserved ◽  
Proliferating Cell

Eukaryotes ubiquitylate the replication factor PCNA (proliferating-cell nuclear antigen) so that it tolerates DNA damage. Although, in the last few years, the understanding of the evolutionarily conserved mechanism of ubiquitylation of PCNA, and its crucial role in DNA damage tolerance, has progressed impressively, little is known about the deubiquitylation of this sliding clamp in most organisms. In the present review, we will discuss potential molecular mechanisms regulating PCNA deubiquitylation in yeast.

scholarly journals The ADP-ribosyltransferase PARP10/ARTD10 Interacts with Proliferating Cell Nuclear Antigen (PCNA) and Is Required for DNA Damage Tolerance

2014 ◽  
Vol 289 (19) ◽  
pp. 13627-13637 ◽  
Author(s):  
Claudia M. Nicolae ◽  
Erin R. Aho ◽  
Alexander H. S. Vlahos ◽  
Katherine N. Choe ◽  
Subhajyoti De ◽  
...  
Keyword(s):  
Dna Damage ◽  
Proliferating Cell Nuclear Antigen ◽  
Damage Tolerance ◽  
Nuclear Antigen ◽  
Dna Damage Tolerance ◽  
Adp Ribosyltransferase ◽  
Proliferating Cell

scholarly journals DNA damage tolerance pathway involving DNA polymerase ι and the tumor suppressor p53 regulates DNA replication fork progression

2016 ◽  
Vol 113 (30) ◽  
pp. E4311-E4319 ◽  
Author(s):  
Stephanie Hampp ◽  
Tina Kiessling ◽  
Kerstin Buechle ◽  
Sabrina F. Mansilla ◽  
Jürgen Thomale ◽  
...  
Keyword(s):  
Dna Damage ◽  
Dna Replication ◽  
Tumor Suppressor ◽  
Damage Tolerance ◽  
Replication Stress ◽  
Nuclear Antigen ◽  
Replication Forks ◽  
Tumor Suppressor P53 ◽  
Dna Damage Tolerance ◽  
Direct Role

DNA damage tolerance facilitates the progression of replication forks that have encountered obstacles on the template strands. It involves either translesion DNA synthesis initiated by proliferating cell nuclear antigen monoubiquitination or less well-characterized fork reversal and template switch mechanisms. Herein, we characterize a novel tolerance pathway requiring the tumor suppressor p53, the translesion polymerase ι (POLι), the ubiquitin ligase Rad5-related helicase-like transcription factor (HLTF), and the SWI/SNF catalytic subunit (SNF2) translocase zinc finger ran-binding domain containing 3 (ZRANB3). This novel p53 activity is lost in the exonuclease-deficient but transcriptionally active p53(H115N) mutant. Wild-type p53, but not p53(H115N), associates with POLι in vivo. Strikingly, the concerted action of p53 and POLι decelerates nascent DNA elongation and promotes HLTF/ZRANB3-dependent recombination during unperturbed DNA replication. Particularly after cross-linker–induced replication stress, p53 and POLι also act together to promote meiotic recombination enzyme 11 (MRE11)-dependent accumulation of (phospho-)replication protein A (RPA)-coated ssDNA. These results implicate a direct role of p53 in the processing of replication forks encountering obstacles on the template strand. Our findings define an unprecedented function of p53 and POLι in the DNA damage response to endogenous or exogenous replication stress.

scholarly journals The Mgs1/WRNIP1 ATPase is required to prevent a recombination salvage pathway at damaged replication forks

2020 ◽  
Vol 6 (15) ◽  
pp. eaaz3327 ◽  
Author(s):  
Alberto Jiménez-Martín ◽  
Irene Saugar ◽  
Chinnu Rose Joseph ◽  
Alexandra Mayer ◽  
Carl P. Lehmann ◽  
...  
Keyword(s):  
Dna Damage ◽  
Genome Stability ◽  
Dna Lesions ◽  
Template Switching ◽  
Replication Forks ◽  
Salvage Pathway ◽  
Dna Damage Tolerance ◽  
Alternative Mode ◽  
Free Mode ◽  
Template Switch

DNA damage tolerance (DDT) is crucial for genome integrity maintenance. DDT is mainly carried out by template switch recombination, an error-free mode of overcoming DNA lesions, or translesion DNA synthesis, which is error-prone. Here, we investigated the role of Mgs1/WRNIP1 in modulating DDT. Using budding yeast, we found that elimination of Mgs1 in cells lacking Rad5, an essential protein for DDT, activates an alternative mode of DNA damage bypass, driven by recombination, which allows chromosome replication and cell viability under stress conditions that block DNA replication forks. This salvage pathway is RAD52 and RAD59 dependent, requires the DNA polymerase δ and PCNA modification at K164, and is enabled by Esc2 and the PCNA unloader Elg1, being inhibited when Mgs1 is present. We propose that Mgs1 is necessary to prevent a potentially toxic recombination salvage pathway at sites of perturbed replication, which, in turn, favors Rad5-dependent template switching, thus helping to preserve genome stability.

scholarly journals Srs2 Plays a Critical Role in Reversible G2 Arrest upon Chronic and Low Doses of UV Irradiation via Two Distinct Homologous Recombination-Dependent Mechanisms in Postreplication Repair-Deficient Cells

2010 ◽  
Vol 30 (20) ◽  
pp. 4840-4850 ◽  
Author(s):  
Takashi Hishida ◽  
Yoshihiro Hirade ◽  
Nami Haruta ◽  
Yoshino Kubota ◽  
Hiroshi Iwasaki
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Excision Repair ◽  
Critical Role ◽  
Nuclear Antigen ◽  
Dna Damage Tolerance ◽  
Checkpoint Activation ◽  
Postreplication Repair ◽  
Nucleotide Excision ◽  
Proliferating Cell

ABSTRACT Differential posttranslational modification of proliferating cell nuclear antigen (PCNA) by ubiquitin or SUMO plays an important role in coordinating the processes of DNA replication and DNA damage tolerance. Previously it was shown that the loss of RAD6-dependent error-free postreplication repair (PRR) results in DNA damage checkpoint-mediated G2 arrest in cells exposed to chronic low-dose UV radiation (CLUV), whereas wild-type and nucleotide excision repair-deficient cells are largely unaffected. In this study, we report that suppression of homologous recombination (HR) in PRR-deficient cells by Srs2 and PCNA sumoylation is required for checkpoint activation and checkpoint maintenance during CLUV irradiation. Cyclin-dependent kinase (CDK1)-dependent phosphorylation of Srs2 did not influence checkpoint-mediated G2 arrest or maintenance in PRR-deficient cells but was critical for HR-dependent checkpoint recovery following release from CLUV exposure. These results indicate that Srs2 plays an important role in checkpoint-mediated reversible G2 arrest in PRR-deficient cells via two separate HR-dependent mechanisms. The first (required to suppress HR during PRR) is regulated by PCNA sumoylation, whereas the second (required for HR-dependent recovery following CLUV exposure) is regulated by CDK1-dependent phosphorylation.

scholarly journals DNA damage-specific deubiquitination regulates Rad18 functions to suppress mutagenesis

2014 ◽  
Vol 206 (2) ◽  
pp. 183-197 ◽  
Author(s):  
Michelle K. Zeman ◽  
Jia-Ren Lin ◽  
Raimundo Freire ◽  
Karlene A. Cimprich
Keyword(s):  
Dna Damage ◽  
Dna Lesions ◽  
Nuclear Antigen ◽  
Lesion Bypass ◽  
Dna Damage Tolerance ◽  
E3 Ligases ◽  
Plant Homeodomain ◽  
In Trans ◽  
Fork Stalling ◽  
Proliferating Cell

Deoxyribonucleic acid (DNA) lesions encountered during replication are often bypassed using DNA damage tolerance (DDT) pathways to avoid prolonged fork stalling and allow for completion of DNA replication. Rad18 is a central E3 ubiquitin ligase in DDT, which exists in a monoubiquitinated (Rad18•Ub) and nonubiquitinated form in human cells. We find that Rad18 is deubiquitinated when cells are treated with methyl methanesulfonate or hydrogen peroxide. The ubiquitinated form of Rad18 does not interact with SNF2 histone linker plant homeodomain RING helicase (SHPRH) or helicase-like transcription factor, two downstream E3 ligases needed to carry out error-free bypass of DNA lesions. Instead, it interacts preferentially with the zinc finger domain of another, nonubiquitinated Rad18 and may inhibit Rad18 function in trans. Ubiquitination also prevents Rad18 from localizing to sites of DNA damage, inducing proliferating cell nuclear antigen monoubiquitination, and suppressing mutagenesis. These data reveal a new role for monoubiquitination in controlling Rad18 function and suggest that damage-specific deubiquitination promotes a switch from Rad18•Ub–Rad18 complexes to the Rad18–SHPRH complexes necessary for error-free lesion bypass in cells.

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