DNA polymerase .epsilon. interacts with proliferating cell nuclear antigen in primer recognition and elongation

Maga, Giovanni; Hübscher, Ulrich

doi:10.1021/bi00003a023

Cited by 37 publications

(35 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This indicates a role for PCNA in the SSA pathway, most likely in the DNA synthesis step following 39 tail removal. Consistent with this idea, PCNA was shown to stabilize binding of various polymerases to the 39-OH of a DNA template during replication and to stimulate DNA synthesis (Maga and Hübscher 1995;Einolf and Guengerich 2000;Maga et al 2002), and human PCNA has been shown to stimulate DNA synthesis during microhomology-mediated end joining, a process similar to SSA that is initiated on terminal microsatellite sequences on ssDNA (Crespan et al 2012). …”

Section: Discussionmentioning

confidence: 78%

A Delicate Balance Between Repair and Replication Factors Regulates Recombination Between Divergent DNA Sequences inSaccharomyces cerevisiae

Chakraborty¹,

George²,

Lyndaker

et al. 2015

Genetics

View full text Add to dashboard Cite

Single-strand annealing (SSA) is an important homologous recombination mechanism that repairs DNA double strand breaks (DSBs) occurring between closely spaced repeat sequences. During SSA, the DSB is acted upon by exonucleases to reveal complementary sequences that anneal and are then repaired through tail clipping, DNA synthesis, and ligation steps. In baker's yeast, the Msh DNA mismatch recognition complex and the Sgs1 helicase act to suppress SSA between divergent sequences by binding to mismatches present in heteroduplex DNA intermediates and triggering a DNA unwinding mechanism known as heteroduplex rejection. Using baker's yeast as a model, we have identified new factors and regulatory steps in heteroduplex rejection during SSA. First we showed that Top3-Rmi1, a topoisomerase complex that interacts with Sgs1, is required for heteroduplex rejection. Second, we found that the replication processivity clamp proliferating cell nuclear antigen (PCNA) is dispensable for heteroduplex rejection, but is important for repairing mismatches formed during SSA. Third, we showed that modest overexpression of Msh6 results in a significant increase in heteroduplex rejection; this increase is due to a compromise in Msh2-Msh3 function required for the clipping of 39 tails. Thus 39 tail clipping during SSA is a critical regulatory step in the repair vs. rejection decision; rejection is favored before the 39 tails are clipped. Unexpectedly, Msh6 overexpression, through interactions with PCNA, disrupted heteroduplex rejection between divergent sequences in another recombination substrate. These observations illustrate the delicate balance that exists between repair and replication factors to optimize genome stability.

show abstract

Section: Discussionmentioning

confidence: 78%

A Delicate Balance Between Repair and Replication Factors Regulates Recombination Between Divergent DNA Sequences inSaccharomyces cerevisiae

Chakraborty¹,

George²,

Lyndaker

et al. 2015

Genetics

View full text Add to dashboard Cite

show abstract

“…Under some conditions, pol e can synthesize DNA e ciently in the absence of PCNA, but under more physiological conditions such as ionic strength greater than 80 mM or the presence of adequate amounts of RPA, pol e needs PCNA to confer signi®cant activity (Lee et al, 1991;Li et al, 1994;Shivji et al, 1995;Maga and HuÈ bscher, 1995). For example, with 145 mM KCl as in Figure 3, ®lling of a short 30 nt gap by both pols e and d is entirely PCNA-dependent.…”

Section: Pcna-dependence Of Nucleotide Excision Repair Synthesismentioning

confidence: 99%

Resistance of human nucleotide excision repair synthesis in vitro to p21Cdn1

et al. 1998

View full text Add to dashboard Cite

The p21 Cdn1 protein (cip1/waf1/sdi1) plays an important role as an inhibitor of mammalian cell proliferation in response to DNA damage. By interacting with and inhibiting the function of cyclin-Cdk complexes, p21 can block entry into S phase. p21 can also directly inhibit replicative DNA synthesis by binding to the DNA polymerase sliding clamp factor PCNA. When cells are damaged and p21 is induced, DNA nucleotide excision repair (NER) continues, even though this pathway is PCNA-dependent. We investigated features of p21-resistant NER using human cell extracts. A direct endlabelling approach was used to measure the excision of damaged oligonucleotides by NER and no inhibition by p21 was found. By contrast, ®lling of the *30 nt gaps created by NER could be inhibited by pre-binding p21 to PCNA, but only when gap ®lling was uncoupled from incision. Binding p21 to PCNA could also inhibit ®lling of model 30 nt gaps by both puri®ed DNA polymerases d and e. When p21 was incubated in a cell extract before addition of PCNA, inhibition of repair synthesis was gradually relieved with time. This incubation gives p21 the opportunity to associate with other targets. As p21 blocks association of DNA polymerases with PCNA but does not prevent loading of PCNA onto DNA, repair gap ®lling can occur rapidly as soon as p21 dissociates from PCNA. A synthetic PCNA-binding p21 peptide was an e cient inhibitor of NER synthesis in cell extracts.

show abstract

“…PCNA, which is encoded by the yeast POL30 gene, serves as an accessory factor for DNA polymerases ␦ and ⑀ and acts by tethering these polymerases to their template DNA during DNA replication, thus enhancing their processivity (22)(23)(24). During DNA replication, PCNA is loaded as a trimer around DNA forming a homotrimeric, sliding, ring-shaped clamp (25,26).…”

mentioning

confidence: 99%

Proliferating Cell Nuclear Antigen (PCNA) Is Required for Cell Cycle-regulated Silent Chromatin on Replicated and Nonreplicated Genes

Miller

Chen

Takasuka

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

In Saccharomyces cerevisiae, silent chromatin is formed at HMR upon the passage through S phase, yet neither the initiation of DNA replication at silencers nor the passage of a replication fork through HMR is required for silencing. Paradoxically, mutations in the DNA replication processivity factor, POL30, disrupt silencing despite this lack of requirement for DNA replication in the establishment of silencing. We tested whether pol30 mutants could establish silencing at either replicated or non-replicated HMR loci during S phase and found that pol30 mutants were defective in establishing silencing at HMR regardless of its replication status. Although previous studies tie the silencing defect of pol30 mutants to the chromatin assembly factors Asf1p and CAF-1, we found pol30 mutants did not exhibit a gross defect in packaging HMR into chromatin. Rather, the pol30 mutants exhibited defects in histone modifications linked to ASF1 and CAF-1-dependent pathways, including SAS-I-and Rtt109p-dependent acetylation events at H4-K16 and H3-K9 (plus H3-K56; Miller, A., Yang, B., Foster, T., and Kirchmaier, A. L. (2008) Genetics 179, 793-809). Additional experiments using FLIM-FRET revealed that Pol30p interacted with SAS-I and Rtt109p in the nuclei of living cells. However, these interactions were disrupted in pol30 mutants with defects linked to ASF1-and CAF-1-dependent pathways. Together, these results imply that Pol30p affects epigenetic processes by influencing the composition of chromosomal histone modifications.To regulate expression of a gene epigenetically, cells must efficiently assemble specialized chromatin at those genes during or shortly after DNA replication each cell cycle. This process ensures that the gene expression state is inherited in future generations. In the budding yeast Saccharomyces cerevisiae, an epigenetic process called silencing prevents transcription of the mating-type genes at the silent mating-type loci HMR and HML (1). When silent chromatin is first formed, the silent information regulator proteins Sir1p, Sir2p, Sir3p, and Sir4p are recruited to the HM loci through their physical interactions with proteins bound to DNA elements called silencers adjacent to the HM loci. At HMR, the four Sir proteins interact with each other and with the origin recognition complex (ORC), 2 Rap1p and Abf1p bound to the E silencer (2-9). Sir2p, Sir3p, and Sir4p then spread along the chromosome and across the mating-type genes a2 and a1 at HMR as the deacetylase Sir2p removes acetyl groups from histones H3 and H4 and creates binding sites for Sir3p and Sir4p on nucleosomes (7, 9) (see also Ref. 5). Once silent chromatin is formed upon passage through S phase, the a2 and a1 genes at HMR are inactivated and their silenced state will be inherited as the genome is duplicated in subsequent generations (1, 10 -14).Although DNA replication itself is not required to establish silent chromatin in S phase (10 -12), a link between DNA replication and silencing in S. cerevisiae has been reinforced by numerous studies. In y...

show abstract

DNA polymerase .epsilon. interacts with proliferating cell nuclear antigen in primer recognition and elongation

Cited by 37 publications

References 46 publications

A Delicate Balance Between Repair and Replication Factors Regulates Recombination Between Divergent DNA Sequences inSaccharomyces cerevisiae

A Delicate Balance Between Repair and Replication Factors Regulates Recombination Between Divergent DNA Sequences inSaccharomyces cerevisiae

Resistance of human nucleotide excision repair synthesis in vitro to p21Cdn1

Proliferating Cell Nuclear Antigen (PCNA) Is Required for Cell Cycle-regulated Silent Chromatin on Replicated and Nonreplicated Genes

Contact Info

Product

Resources

About