scholarly journals Rad27 and Exo1 function in different excision pathways for mismatch repair in Saccharomyces cerevisiae

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Felipe A. Calil ◽  
Bin-Zhong Li ◽  
Kendall A. Torres ◽  
Katarina Nguyen ◽  
Nikki Bowen ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Dna Mismatch Repair ◽  
Dna Mismatch ◽  
Eukaryotic Dna ◽  
Mutl Homolog 1 ◽  
Exonuclease 1 ◽  
Dna Substrate ◽  
Mmr Proteins

AbstractEukaryotic DNA Mismatch Repair (MMR) involves redundant exonuclease 1 (Exo1)-dependent and Exo1-independent pathways, of which the Exo1-independent pathway(s) is not well understood. The exo1Δ440-702 mutation, which deletes the MutS Homolog 2 (Msh2) and MutL Homolog 1 (Mlh1) interacting peptides (SHIP and MIP boxes, respectively), eliminates the Exo1 MMR functions but is not lethal in combination with rad27Δ mutations. Analyzing the effect of different combinations of the exo1Δ440-702 mutation, a rad27Δ mutation and the pms1-A99V mutation, which inactivates an Exo1-independent MMR pathway, demonstrated that each of these mutations inactivates a different MMR pathway. Furthermore, it was possible to reconstitute a Rad27- and Msh2-Msh6-dependent MMR reaction in vitro using a mispaired DNA substrate and other MMR proteins. Our results demonstrate Rad27 defines an Exo1-independent eukaryotic MMR pathway that is redundant with at least two other MMR pathways.

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

2017 ◽  
Vol 114 (14) ◽  
pp. 3607-3612 ◽  
Author(s):  
Nikki Bowen ◽  
Richard D. Kolodner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Dna Polymerase ◽  
Strand Break ◽  
Single Strand Break ◽  
Mutl Homolog 1 ◽  
Exonuclease 1 ◽  
Postmeiotic Segregation ◽  
Mmr Proteins

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 ε (Pol ε) may also play a role in MMR. Here we describe the reconstitution of Pol ε-dependent MMR using S. cerevisiae proteins. A mixture of Msh2–Msh6 (or Msh2–Msh3), Exo1, RPA, RFC-Δ1N, PCNA, and Pol ε 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 ε was found to catalyze an MMR reaction that required Mlh1–Pms1. These results demonstrate that Pol ε can act in eukaryotic MMR in vitro.

Heteroduplex DNA correction in Saccharomyces cerevisiae is mismatch specific and requires functional PMS genes

1989 ◽  
Vol 9 (10) ◽  
pp. 4432-4440
Author(s):  
B Kramer ◽  
W Kramer ◽  
M S Williamson ◽  
S Fogel
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Dna Mismatch Repair ◽  
Evolutionary Relationship ◽  
Close Correlation ◽  
Single Nucleotide ◽  
Dna Mismatch ◽  
Low Efficiency ◽  
Specificity Pattern

In vitro-constructed heteroduplex DNAs with defined mismatches were corrected in Saccharomyces cerevisiae cells with efficiencies that were dependent on the mismatch. Single-nucleotide loops were repaired very efficiently; the base/base mismatches G/T, A/C, G/G, A/G, G/A, A/A, T/T, T/C, and C/T were repaired with a high to intermediate efficiency. The mismatch C/C and a 38-nucleotide loop were corrected with low efficiency. This substrate specificity pattern resembles that found in Escherichia coli and Streptococcus pneumoniae, suggesting an evolutionary relationship of DNA mismatch repair in pro- and eucaryotes. Repair of the listed mismatches was severely impaired in the putative S. cerevisiae DNA mismatch repair mutants pms1 and pms2. Low-efficiency repair also characterized pms3 strains, except that correction of single-nucleotide loops occurred with an efficiency close to that of PMS wild-type strains. A close correlation was found between the repair efficiencies determined in this study and the observed postmeiotic segregation frequencies of alleles with known DNA sequence. This suggests an involvement of DNA mismatch repair in recombination and gene conversion in S. cerevisiae.

scholarly journals Heteroduplex DNA correction in Saccharomyces cerevisiae is mismatch specific and requires functional PMS genes.

1989 ◽  
Vol 9 (10) ◽  
pp. 4432-4440 ◽  
Author(s):  
B Kramer ◽  
W Kramer ◽  
M S Williamson ◽  
S Fogel
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Dna Mismatch Repair ◽  
Evolutionary Relationship ◽  
Close Correlation ◽  
Single Nucleotide ◽  
Dna Mismatch ◽  
Low Efficiency ◽  
Specificity Pattern

In vitro-constructed heteroduplex DNAs with defined mismatches were corrected in Saccharomyces cerevisiae cells with efficiencies that were dependent on the mismatch. Single-nucleotide loops were repaired very efficiently; the base/base mismatches G/T, A/C, G/G, A/G, G/A, A/A, T/T, T/C, and C/T were repaired with a high to intermediate efficiency. The mismatch C/C and a 38-nucleotide loop were corrected with low efficiency. This substrate specificity pattern resembles that found in Escherichia coli and Streptococcus pneumoniae, suggesting an evolutionary relationship of DNA mismatch repair in pro- and eucaryotes. Repair of the listed mismatches was severely impaired in the putative S. cerevisiae DNA mismatch repair mutants pms1 and pms2. Low-efficiency repair also characterized pms3 strains, except that correction of single-nucleotide loops occurred with an efficiency close to that of PMS wild-type strains. A close correlation was found between the repair efficiencies determined in this study and the observed postmeiotic segregation frequencies of alleles with known DNA sequence. This suggests an involvement of DNA mismatch repair in recombination and gene conversion in S. cerevisiae.

Eukaryotic DNA Mismatch Repair In Vitro

2012 ◽  
pp. 149-162 ◽  
Author(s):  
Fenghua Yuan ◽  
Limin Song ◽  
Fengsong Liu ◽  
Liya Gu ◽  
Yanbin Zhang
Keyword(s):  
Mismatch Repair ◽  
Dna Mismatch Repair ◽  
Dna Mismatch ◽  
Eukaryotic Dna

scholarly journals Nuclear localization of human DNA mismatch repair protein exonuclease 1 (hEXO1)

2007 ◽  
Vol 35 (8) ◽  
pp. 2609-2619 ◽  
Author(s):  
Nina Østergaard Knudsen ◽  
Finn Cilius Nielsen ◽  
Lena Vinther ◽  
Ronni Bertelsen ◽  
Steen Holten-Andersen ◽  
...  
Keyword(s):  
Mismatch Repair ◽  
Nuclear Localization ◽  
Dna Mismatch Repair ◽  
Dna Mismatch ◽  
Human Dna ◽  
Repair Protein ◽  
Exonuclease 1 ◽  
Mismatch Repair Protein

scholarly journals Functional Studies on the Candidate ATPase Domains of Saccharomyces cerevisiae MutLα

2000 ◽  
Vol 20 (17) ◽  
pp. 6390-6398 ◽  
Author(s):  
Phuoc T. Tran ◽  
R. Michael Liskay
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Conformational Changes ◽  
Atp Hydrolysis ◽  
Dna Mismatch Repair ◽  
Limited Proteolysis ◽  
Atp Binding ◽  
Dna Mismatch ◽  
Functional Studies

ABSTRACT Saccharomyces cerevisiae MutL homologues Mlh1p and Pms1p form a heterodimer, termed MutLα, that is required for DNA mismatch repair after mismatch binding by MutS homologues. Recent sequence and structural studies have placed the NH2 termini of MutL homologues in a new family of ATPases. To address the functional significance of this putative ATPase activity in MutLα, we mutated conserved motifs for ATP hydrolysis and ATP binding in both Mlh1p and Pms1p and found that these changes disrupted DNA mismatch repair in vivo. Limited proteolysis with purified recombinant MutLα demonstrated that the NH2 terminus of MutLα undergoes conformational changes in the presence of ATP and nonhydrolyzable ATP analogs. Furthermore, two-hybrid analysis suggested that these ATP-binding-induced conformational changes promote an interaction between the NH2 termini of Mlh1p and Pms1p. Surprisingly, analysis of specific mutants suggested differential requirements for the ATPase motifs of Mlh1p and Pms1p during DNA mismatch repair. Taken together, these results suggest that MutLα undergoes ATP-dependent conformational changes that may serve to coordinate downstream events during yeast DNA mismatch repair.

scholarly journals Involvement of DNA mismatch repair in the maintenance of heterochromatic DNA stability in Saccharomyces cerevisiae

PLoS Genetics ◽  
2017 ◽  
Vol 13 (10) ◽  
pp. e1007074 ◽  
Author(s):  
Basanta K. Dahal ◽  
Lyudmila Y. Kadyrova ◽  
Kristin R. Delfino ◽  
Igor B. Rogozin ◽  
Vaibhavi Gujar ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mismatch Repair ◽  
Dna Mismatch Repair ◽  
Dna Stability ◽  
Dna Mismatch
Sign in / Sign up

Export Citation Format

Share Document