Mismatch Repair, But Not Heteroduplex Rejection, Is Temporally Coupled to DNA Replication

Hombauer, Hans; Srivatsan, Anjana; Putnam, Christopher D.; Kolodner, Richard D.

doi:10.1126/science.1210770

Cited by 112 publications

(119 citation statements)

References 27 publications

(36 reference statements)

Supporting

Mentioning

115

Contrasting

Order By: Relevance

“…Finally, the strong enrichment of the mismatch repair proteins MSH2, MSH3, and MSH6 at active elongating forks is consistent with recent data from yeast systems indicating that these proteins travel with the replisome (29). We verified that both MSH2 and MSH6 are associated with the replisome in a pattern mirroring PCNA using conventional immunoblotting (data not shown).…”

Section: ϫ11supporting

confidence: 75%

Identification of Proteins at Active, Stalled, and Collapsed Replication Forks Using Isolation of Proteins on Nascent DNA (iPOND) Coupled with Mass Spectrometry

Sirbu¹,

McDonald

Dungrawala³

et al. 2013

Journal of Biological Chemistry

219

226

View full text Add to dashboard Cite

Background: DNA replication and the replication stress response require the coordinated actions of many proteins. Results: iPOND coupled with mass spectrometry identified 290 proteins associated with active, stalled, or collapsed replication forks. Conclusion: iPOND-MS is a useful discovery tool. Significance: The data increase our understanding of the network of proteins involved in DNA replication and the replication stress response.

show abstract

Section: ϫ11supporting

confidence: 75%

Identification of Proteins at Active, Stalled, and Collapsed Replication Forks Using Isolation of Proteins on Nascent DNA (iPOND) Coupled with Mass Spectrometry

Sirbu¹,

McDonald

Dungrawala³

et al. 2013

Journal of Biological Chemistry

219

226

View full text Add to dashboard Cite

show abstract

“…We recently showed that MMR is coupled to DNA replication and that the mispair recognition proteins Msh2-Msh6 and Msh2-Msh3 colocalize with the replicative DNA polymerases during DNA replication (55,56). This latter observation led us to investigate whether Pol e might also function in MMR in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…MMR appears to be coupled to the replication fork as evidenced by the timing of MMR and the colocalization of the Msh2-Msh6 mispair recognition complex with replication fork proteins during S phase (55,56). Previous studies suggesting that only Pol δ acts in MMR (50) could be interpreted as implying that the selection of the DNA polymerase used for the gap-filling step of MMR is not directly coupled to DNA replication because Pol δ does not act in leading-strand DNA synthesis.…”

Section: Discussionmentioning

confidence: 99%

Reconstitution of Saccharomyces cerevisiae DNA polymerase ε-dependent mismatch repair with purified proteins

Bowen

Kolodner

2017

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Mammalian and Saccharomyces cerevisiae mismatch repair (MMR) proteins catalyze two MMR reactions in vitro. In one, mispair binding by either the MutS homolog 2 (Msh2)-MutS homolog 6 (Msh6) or the Msh2-MutS homolog 3 (Msh3) stimulates 5′ to 3′ excision by exonuclease 1 (Exo1) from a single-strand break 5′ to the mispair, excising the mispair. In the other, Msh2-Msh6 or Msh2-Msh3 activate the MutL homolog 1 (Mlh1)-postmeiotic segregation 1 (Pms1) endonuclease in the presence of a mispair and a nick 3′ to the mispair, to make nicks 5′ to the mispair, allowing Exo1 to excise the mispair. DNA polymerase δ (Pol δ) is thought to catalyze DNA synthesis to fill in the gaps resulting from mispair excision. However, colocalization of the S. cerevisiae mispair recognition proteins with the replicative DNA polymerases during DNA replication has suggested that DNA polymerase e (Pol e) may also play a role in MMR. Here we describe the reconstitution of Pol e-dependent MMR using S. cerevisiae proteins. A mixture of Msh2-Msh6 (or Msh2-Msh3), Exo1, RPA, RFC-Δ1N, PCNA, and Pol e was found to catalyze both short-patch and long-patch 5′ nick-directed MMR of a substrate containing a +1 (+T) mispair. When the substrate contained a nick 3′ to the mispair, a mixture of Msh2-Msh6 (or Msh2-Msh3), Exo1, RPA, RFC-Δ1N, PCNA, and Pol e was found to catalyze an MMR reaction that required Mlh1-Pms1. These results demonstrate that Pol e can act in eukaryotic MMR in vitro.mutator phenotype | genome instability | DNA replication fidelity | DNA excision | DNA repair

show abstract

“…Studies of bacterial MMR have provided a basic mechanistic framework for comparative studies (5). Genetic and cell-biology studies, primarily in Saccharomyces cerevisiae, have identified eukaryotic MMR genes, provided models for how their gene products define MMR pathways, and elucidated some of the details of how MMR pathways interact with replication (1)(2)(3)(4). Reconstitution studies, primarily in human systems, have identified some of the catalytic features of eukaryotic MMR (7-9, 16, 17).…”

mentioning

confidence: 99%

“…DNA replication fidelity | genome instability | mutator phenotype | cancer | mutagenesis D NA mismatch repair (MMR) is a critical DNA repair pathway that is coupled to DNA replication in eukaryotes where it corrects misincorporation errors made during DNA replication (1)(2)(3)(4)(5)(6)(7)(8)(9). This pathway prevents mutations and acts to prevent the development of cancer (10,11).…”

mentioning

confidence: 99%

Reconstitution of long and short patch mismatch repair reactions using Saccharomyces cerevisiae proteins

Bowen¹,

Smith²,

Srivatsan³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

A problem in understanding eukaryotic DNA mismatch repair (MMR) mechanisms is linking insights into MMR mechanisms from genetics and cell-biology studies with those from biochemical studies of MMR proteins and reconstituted MMR reactions. This type of analysis has proven difficult because reconstitution approaches have been most successful for human MMR whereas analysis of MMR in vivo has been most advanced in the yeast Saccharomyces cerevisiae. Here, we describe the reconstitution of MMR reactions using purified S. cerevisiae proteins and mispaircontaining DNA substrates. A mixture of MutS homolog 2 (Msh2)-MutS homolog 6, Exonuclease 1, replication protein A, replication factor C-Δ1N, proliferating cell nuclear antigen and DNA polymerase δ was found to repair substrates containing TG, CC, +1 (+T), +2 (+GC), and +4 (+ACGA) mispairs and either a 5′ or 3′ strand interruption with different efficiencies. The Msh2-MutS homolog 3 mispair recognition protein could substitute for the Msh2-Msh6 mispair recognition protein and showed a different specificity of repair of the different mispairs whereas addition of MutL homolog 1-postmeiotic segregation 1 had no affect on MMR. Repair was catalytic, with as many as 11 substrates repaired per molecule of Exo1. Repair of the substrates containing either a 5′ or 3′ strand interruption occurred by mispair binding-dependent 5′ excision and subsequent resynthesis with excision tracts of up to ∼2.9 kb occurring during the repair of the substrate with a 3′ strand interruption. The availability of this reconstituted MMR reaction now makes possible detailed biochemical studies of the wealth of mutations identified that affect S. cerevisiae MMR.DNA replication fidelity | genome instability | mutator phenotype | cancer | mutagenesis

show abstract

Mismatch Repair, But Not Heteroduplex Rejection, Is Temporally Coupled to DNA Replication

Cited by 112 publications

References 27 publications

Identification of Proteins at Active, Stalled, and Collapsed Replication Forks Using Isolation of Proteins on Nascent DNA (iPOND) Coupled with Mass Spectrometry

Identification of Proteins at Active, Stalled, and Collapsed Replication Forks Using Isolation of Proteins on Nascent DNA (iPOND) Coupled with Mass Spectrometry

Reconstitution of Saccharomyces cerevisiae DNA polymerase ε-dependent mismatch repair with purified proteins

Reconstitution of long and short patch mismatch repair reactions using Saccharomyces cerevisiae proteins

Contact Info

Product

Resources

About