The cell cycle and DNA mismatch repair

Schroering, Allen; Edelbrock, Michael A.; Richards, Timothy J.; Williams, Kandace J.

doi:10.1016/j.yexcr.2006.10.018

Cited by 56 publications

(89 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The dramatic increase of detergentresistant MMR proteins 2 hours after MNNG treatment, even in the absence of DNA synthesis, agrees well with our ChIP results indicating rapid recruitment and co-localized binding to the alkylation-damaged chromatin. Interestingly, ChIP analysis ( Figure 2a) and microscopic immunofluorescence (Supplement 3) of untreated cells in S phase indicates a constititutive level of MutSα associated with the chromatin which is also localized to discrete replication factories during S phase, as previously reported by us and others [37,41]. However, detergent extraction before immunofluorescence staining indicates that MutSα is extractable, and therefore not actually bound to undamaged chromosomal DNA that has not been subjected to formaldehyde cross-linking (Figure 3a; 0 MNNG).…”

Section: Effects Of Cell Cycle Synchronization Dna Replication Blocksupporting

confidence: 83%

“…Within the nucleus of both untreated HeLa S3 and MR cells synchronized to S phase, MutSα is primarily localized to discrete replication factories, as previously reported (Supplement 3) [37,41,42]. Two hours after exposure to 20 µM MNNG however, a dramatic increase in nuclear accumulation of MSH6 is evident, and the pattern of distribution in both HeLa S3 and MR cells is much more diffuse throughout the nucleus (Supplement 3).…”

Section: Effects Of Cell Cycle Synchronization Dna Replication Blocksupporting

confidence: 79%

“…Note that the majority of the G 1 synchronized cells have just begun to enter S phase by 4 hours and 70% of the population is still in S phase by 8 hours (Figure 5a, b). Therefore, soluble MMR protein concentrations within the nucleus of alkylation-damaged cells may also be influenced by the cell cycle as well as MNNG treatment [37]. In summary, we have demonstrated that increased chromatin association of MMR proteins in HeLa MR cells can be rapidly induced by exposure to MNNG in a concentration-dependent manner, does not require DNA replication, and is a separate induction from the normal cell cycle fluctuation of nuclear MMR protein concentrations [37].…”

Section: Effects Of Cell Cycle Synchronization Dna Replication Blockmentioning

confidence: 82%

“…Monitoring of cell cycle phase distribution was accomplished by using propidium iodide for nuclear staining and subsequent detection using a Beckman/Coulter EPICS Elite flow cytometer, as described previously [37]. The resulting data were analyzed by Multicycle software (Phoenix Flow Systems) and reported as the percentage of cells in G 1 , S, or G 2 phase.…”

Section: Cell Cycle Analysesmentioning

confidence: 99%

See 3 more Smart Citations

Rapid induction of chromatin-associated DNA mismatch repair proteins after MNNG treatment

Schroering

Williams

2008

DNA Repair

Self Cite

View full text Add to dashboard Cite

Treatment with low concentrations of monofunctional alkylating agents induces a G 2 arrest only after the second round of DNA synthesis in mammalian cells and requires a proficient mismatch repair (MMR) pathway. Here we have investigated rapid alkylation-induced recruitment of DNA repair proteins to chromosomal DNA within synchronized populations of MMR proficient cells (HeLa MR) after MNNG treatment. Within the first hour, the concentrations of MutSα and PCNA increase well beyond their constitutive chromosomally bound levels and MutLα is newly recruited to the chromatin-bound MutSα. Remarkably, immunoprecipitation experiments demonstrate rapid association of these proteins on the alkylation-damaged chromatin, even when DNA replication is completely blocked. The extent of association of PCNA and MMR proteins on the chromatin is dependent upon the concentration of MNNG and on the specific type of replication block. A subpopulation of the MutSα-associated PCNA also becomes monoubiquitinated, a known requirement for PCNA to interact with translesion synthesis (TLS) polymerases. In addition, chromatin-bound SMC1 and NBS1 proteins, associated with DNA double-strand-breaks (DSBs), become phosphorylated within one to two hours of exposure to MNNG. However, these activated proteins are not colocalized on the chromatin with MutSα in response to MNNG exposure. PCNA, MutSα/MutLα and activated SMC1/NBS1 remain chromatin-bound for at least 6-8 hours after alkylation damage. Thus, cells that are exposed to low levels of alkylation treatment undergo rapid recruitment to and/or activation of key proteins already on the chromatin without the requirement for DNA replication, apparently via different DNA-damage signaling pathways.

show abstract

Section: Effects Of Cell Cycle Synchronization Dna Replication Blocksupporting

confidence: 83%

Section: Effects Of Cell Cycle Synchronization Dna Replication Blocksupporting

confidence: 79%

Section: Effects Of Cell Cycle Synchronization Dna Replication Blockmentioning

confidence: 82%

Section: Cell Cycle Analysesmentioning

confidence: 99%

See 2 more Smart Citations

Rapid induction of chromatin-associated DNA mismatch repair proteins after MNNG treatment

Schroering

Williams

2008

DNA Repair

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, others have suggested that resting intestinal cells did not express MSH2 (69) and the MMR protein levels exhibited fluctuations (70). Another recent investigation has determined that the expression levels of MMR mRNA and protein are up-regulated by growth stimulation (71,72). In this study, we found that MSH2 expression level and MMR activity were decreased in quiescent cells.…”

Section: Discussionsupporting

confidence: 53%

Senescence-Dependent MutSα Dysfunction Attenuates Mismatch Repair

Chang¹,

Jin

Yoon

et al. 2008

Molecular Cancer Research

View full text Add to dashboard Cite

DNA damage and mutations in the genome increase with age. To determine the potential mechanisms of senescence-dependent increases in genomic instability, we analyzed DNA mismatch repair (MMR) efficiency in young and senescent human colonic fibroblast and human embryonic lung fibroblast. It was found that MMR activity is significantly reduced in senescent cells. Western blot and immunohistochemistry analysis revealed that hMSH2 and MSH6 protein (MutSA complex), which is a known key component in the MMR pathway, is markedly down-regulated in senescent cells. Moreover, the addition of purified MutSA to extracts from senescent cells led to the restoration of MMR activity. Semiquantitative reverse transcription-PCR analysis exhibited that MSH2 mRNA level is reduced in senescent cells. In addition, a decrease in E2F transcriptional activity in senescent cells was found to be crucial for MSH2 suppression. E2F1 small interfering RNA expression reduced hMSH2 expression and MMR activity in young human primary fibroblast cells. Importantly, expression of E2F1 in quiescent cells restored the MSH2 expression as well as MMR activity, whereas E2F1-infected senescent cells exhibited no restoration of MSH2 expression and MMR activity. These results indicate that the suppression of E2F1 transcriptional activity in senescent cells lead to stable repression of MSH2, followed by a induction of MutSA dysfunction, which results in a reduced cellular MMR capacity in senescent cells. (Mol Cancer Res 2008;6(6):978 -89)

show abstract

Human DNA polymerase α interacts with mismatch repair proteins MSH2 and MSH6

et al. 2016

View full text Add to dashboard Cite

High fidelity of genome duplication is ensured by cooperation of polymerase proofreading and mismatch repair (MMR) activities. Here, we show that human mismatch recognizing proteins MutS homolog 2 (MSH2) and MSH6 copurify and interact with replicative Pol α. This enzyme also is the replicative primase and replicates DNA with poor fidelity. We show that MSH2 associates with known human replication origins with different dynamics than DNA polymerase (Pol α). Furthermore, we explored the potential functional role of Pol α in the mismatch repair reaction using an in vitro mismatch repair assay and observed that Pol α promotes mismatch repair. Taken together, we show that human Pol α interacts with MSH2-MSH6 complex and propose that this interaction occurs during the mismatch repair reaction.

show abstract

The cell cycle and DNA mismatch repair

Cited by 56 publications

References 35 publications

Rapid induction of chromatin-associated DNA mismatch repair proteins after MNNG treatment

Rapid induction of chromatin-associated DNA mismatch repair proteins after MNNG treatment

Senescence-Dependent MutSα Dysfunction Attenuates Mismatch Repair

Human DNA polymerase α interacts with mismatch repair proteins MSH2 and MSH6

Contact Info

Product

Resources

About