Ligation of newly replicated DNA controls the timing of DNA mismatch repair

Reyes, Gloria; Kolodziejczak, Anna; Devakumar, Lovely Jael Paul Solomon; Kubota, Takashi; Kolodner, Richard D.; Putnam, Christopher D.; Hombauer, Hans

doi:10.1016/j.cub.2020.12.018

Cited by 22 publications

(20 citation statements)

References 55 publications

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“…As for E. coli MutL foci, long-lived PMS1 and MHL2 foci could mark unrepairable mismatches converted into mutations [ 163 ]. An increased frequency of PMS1 foci correlating with the mutator phenotype of the cells, potentially due to an increased PMS1 foci lifespan, was also recently reported in an exo1 strain overproducing CDC9 DNA ligase [ 164 ]. This study suggests that ligation of newly replicated DNA controls the timing of MMR in S. cerevisiae and possibly other eukaryotes.…”

Section: Visualization Of Mmr Proteins In Live Cellsmentioning

confidence: 64%

Mismatch Repair: From Preserving Genome Stability to Enabling Mutation Studies in Real-Time Single Cells

Elez

2021

Cells

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Mismatch Repair (MMR) is an important and conserved keeper of the maintenance of genetic information. Miroslav Radman’s contributions to the field of MMR are multiple and tremendous. One of the most notable was to provide, along with Bob Wagner and Matthew Meselson, the first direct evidence for the existence of the methyl-directed MMR. The purpose of this review is to outline several aspects and biological implications of MMR that his work has helped unveil, including the role of MMR during replication and recombination editing, and the current understanding of its mechanism. The review also summarizes recent discoveries related to the visualization of MMR components and discusses how it has helped shape our understanding of the coupling of mismatch recognition to replication. Finally, the author explains how visualization of MMR components has paved the way to the study of spontaneous mutations in living cells in real time.

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Section: Visualization Of Mmr Proteins In Live Cellsmentioning

confidence: 64%

Mismatch Repair: From Preserving Genome Stability to Enabling Mutation Studies in Real-Time Single Cells

Elez

2021

Cells

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“…The protocol used for chromatin enrichment is described in (70). Around 40 OD cells were harvested from a logarithmically growing yeast culture and resuspended in 1 mL of pre-spheroplasting buffer (100 mM PIPES/KOH, pH 9.4, 10 mM DTT, 0.1% sodium azide).…”

Section: Methodsmentioning

confidence: 99%

“…Next, cells were suspended in spheroplasting buffer (50 mM KH 2 PO 4 /K 2 HPO 4 , pH 7.4, 0.8 M sorbitol, 10 mM DTT, 0.1% sodium azide) containing 200 μg/ml Zymolyase-100T at 30°C for 30 mins on a roller at slow speed. The spheroplasts were confirmed microscopically, and protocol from (70) was followed afterward. The Histone H3 and Rps6 were used as a control for chromatin enrichment.…”

Section: Methodsmentioning

confidence: 99%

S. cerevisiae cells can grow without the Pds5 cohesin subunit

Choudhary

Itzkovich

Alonso-Pérez

et al. 2022

Preprint

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During DNA replication, the newly created sister chromatids are held together until their separation at anaphase. The cohesin complex is in charge of creating and maintaining sister-chromatid cohesion (SCC) in all eukaryotes. In S. cerevisiae cells, cohesin is composed of two elongated proteins, Smc1 and Smc3, bridged by the kleisin Mcd1/Scc1. The latter also acts as a scaffold for three additional proteins, Scc3/Irr1, Wpl1/Rad61, and Pds5. Although the HEAT-repeat protein Pds5 is essential for cohesion, its precise function is still debated. Deletion of the ELG1 gene, encoding a PCNA unloader, can partially suppress the temperature-sensitive pds5-1 allele, but not a complete deletion of PDS5. We carried out a genetic screen for high copy number suppressors and another for spontaneously arising mutants, allowing the survival of a pds5Δ elg1Δ strain. Our results show that cells remain viable in the absence of Pds5 provided that there is both an elevation in the level of Mcd1 (which can be due to mutations in the CLN2 gene, encoding a G1 cyclin), and an increase in the level of SUMO-modified PCNA on chromatin (caused by lack of PCNA unloading in elg1Δ mutants). The elevated SUMO-PCNA levels increase the recruitment of the Srs2 helicase, which evicts Rad51 molecules from the moving fork, creating ssDNA regions that serve as sites for increased cohesin loading and SCC establishment. Thus, our results delineate a double role for Pds5 in protecting the cohesin ring and interacting with the DNA replication machinery.IMPORTANCESister chromatid cohesion is vital for faithful chromosome segregation, chromosome folding into loops, and gene expression. A multisubunit protein complex known as cohesin holds the sister chromatids from S-phase until the anaphase stage. In this study, we explore the function of the essential cohesin subunit Pds5 in the regulation of sister chromatid cohesion. We performed two independent genetic screens to bypass the function of the Pds5 protein. We observe that Pds5 protein is a cohesin stabilizer, and elevating the levels of Mcd1 protein along with SUMO-PCNA accumulation on chromatin can compensate for the loss of the PDS5 gene. In addition, Pds5 plays a role in coordinating the DNA replication and sister chromatid cohesion establishment. This work elucidates the function of cohesin subunit Pds5, the G1 cyclin Cln2, and replication factors PCNA, Elg1 and Srs2 in the proper regulation of sister chromatid cohesion.

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“…The correction of DNA replication errors by the MMR system increases the replication fidelity by ~100 fold (25). Strand breaks in leading and lagging strands as well as ribonucleotides J o u r n a l P r e -p r o o f in leading strands serve as signals that direct the eukaryotic MMR system to remove DNA replication errors (26)(27)(28)(29)(30). MMR is more efficient on the lagging than the leading strand (31).…”

Section: Introductionmentioning

confidence: 99%

The nuclease activity of DNA2 promotes exonuclease 1–independent mismatch repair

Kadyrova

Dahal

Gujar

et al. 2022

Journal of Biological Chemistry

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Ligation of newly replicated DNA controls the timing of DNA mismatch repair

Abstract: Highlights d Cdc9 overexpression causes increased mutation rates and accumulation of Pms1 foci d DNA replication-associated nicks are required for MMR strand discrimination d Cdc9 activity dictates a temporal window for MMR

Cited by 22 publications

References 55 publications

Mismatch Repair: From Preserving Genome Stability to Enabling Mutation Studies in Real-Time Single Cells

Mismatch Repair: From Preserving Genome Stability to Enabling Mutation Studies in Real-Time Single Cells

S. cerevisiae cells can grow without the Pds5 cohesin subunit

The nuclease activity of DNA2 promotes exonuclease 1–independent mismatch repair

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