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Flap endonuclease 1 ( FEN1) and Dna2 are responsible for flap cleavage. Paper-13677576.
SGS1 encodes a helicase with similar properties to Dna2 protein. Paper-8463768.
This gene is identical to DNA2, encoding a helicase required for DNA replication. Paper-8260933.
Okazaki fragment maturation in yeast. I. Distribution of functions between FEN1 AND DNA2. Paper-9868443.
Sgs1 helicase and two nucleases Dna2 and Exo1 resect DNA double-strand break ends. Paper-12950376.
Evidence that yeast SGS1, DNA2, SRS2, and FOB1 interact to maintain rDNA stability. Paper-10042636.
Dna2 tracked onto these flaps but could not cleave, presenting a block to FEN1 entry. Paper-13677576.
DNA2 encodes a DNA helicase essential for replication of eukaryotic chromosomes. Paper-416672.
Specifically, we show that deletion of PIF1 suppresses the lethality of a DNA2-null mutant. Paper-11348595.
Upon cleavage, Dna2 leaves a short flap, which is then cut by FEN1 forming a nick for ligation. Paper-13677576.
However, maturation of long 5'-flaps to which replication protein A can bind required both DNA2 and FEN1. Paper-9868443.
We have recently described a new helicase, the Dna2 helicase, that is essential for yeast DNA replication. Paper-958038.
Previously, we demonstrated that FEN1 disengages the tracking mechanism of Dna2 to remove it from the flap. Paper-13677576.
Coupling of DNA helicase and endonuclease activities of yeast Dna2 facilitates Okazaki fragment processing. Paper-9166542.
Maturation was efficient in the absence of Dna2 and required Dna2 only when FEN1 activity was compromised. Paper-9868442.
Characterization of Saccharomyces cerevisiae dna2 mutants suggests a role for the helicase late in S phase. Paper-8260933.
Among them we found RAD27 and subsequently DNA2 which encode for proteins involved in DNA repair and replication. Paper-12881956.
A second nuclease, Dna2p, is needed to cleave an RPA-coated flap producing a short RPA-free flap, favored by FEN1. Paper-10561072.
Dna2p helicase/nuclease is a tracking protein, like FEN1, for flap cleavage during Okazaki fragment maturation. Paper-10561072.
Here we show that the endonucleases Dna2 and Fen1 act sequentially to facilitate the complete removal of the primer RNA. Paper-9031808.
We propose that a bimodal interaction of Dna2 with Rpa1 is important for Dna2 function both in vivo and in vitro. Paper-9718988.
However, FEN1 disengaged these nonproductively bound Dna2 molecules, proceeding on to conduct proper cleavage. Paper-13677576.
Unlike FEN1, Dna2p shows evidence of a "threading-like" mechanism that does not support tracking over a branched substrate. Paper-10561072.
The Dna2 nuclease/ helicase alone did not efficiently promote nick translation, nor did it affect nick translation with FEN1. Paper-9868443.
We propose that the two nucleases both track, Dna2p first and then FEN1, to remove initiator RNA via long flap intermediates. Paper-10561072.
A yeast replicative helicase, Dna2 helicase, interacts with yeast FEN-1 nuclease in carrying out its essential function. Paper-958038.
Dna2Pho has both a RecB-like nuclease motif and seven conserved helicase motifs similar to Dna2 from Saccharomyces cerevisiae. Paper-9695890.
Like FEN1, Dna2p can track over substrates with a non-Watson Crick base, such as a biotin, or a missing base within a chain. Paper-10561072.
Evidence suggesting that Pif1 helicase functions in DNA replication with the Dna2 helicase/nuclease and DNA polymerase delta. Paper-11348595.
FEN1 cleaves 10-nucleotide fixed or equilibrating flaps in an efficient reaction, insensitive to even high levels of RPA or Dna2p. Paper-10238035.
We show that the Mre11-Rad50-Xrs2 complex (MRX) initiates 5' degradation, whereas Sgs1 and Dna2 degrade 5' strands exposing long 3' strands. Paper-12950376.
Helicase and nuclease activities of hyperthermophile Pyrococcus horikoshii Dna2 inhibited by substrates with RNA segments at 5'-end. Paper-9695890.
Single Strand Annealing and ATP-independent Strand Exchange Activities of Yeast and Human DNA2: POSSIBLE ROLE IN OKAZAKI FRAGMENT MATURATION. Paper-12350212.
The repair genes RAD27 and DNA2 stabilise human minisatellites in yeast mitosis, while RAD5 has no effect on mitotic stability. Paper-12036266.
Downstream DNA primers, RNA primers, and small 5'-flaps were efficiently matured by Pol delta and FEN1, and Dna2 did not stimulate maturation. Paper-9868443.
The Saccharomyces cerevisiae DNA2 gene encodes a DNA- stimulated ATPase and DNA helicase/nuclease essential for DNA replication. Paper-8463768.
However, Dna2p has a role in a pathway for processing structured flaps, in which it aids FEN1 using both its nuclease and helicase activities. Paper-10238035.
The Dna2 protein is a multifunctional enzyme with 5'-3' DNA helicase, DNA-dependent ATPase, 3' exo/endonuclease, and 5' exo/endonuclease. Paper-12350212.
Full activity of the Dna2 helicase function is therefore not needed for viability, but is required for repairing damage and for tolerating loss of Ctf4. Paper-1823220.
Class 2 includes genes involved in the processing of Okazaki fragments in lagging strand synthesis and is represented by SLM1, which is allelic to DNA2. Paper-10017085.
Thus, suppression of dna2delta can be rationalized if deletion of POL32 and/or PIF1 results in a reduction in long flaps that require Dna2 for processing. Paper-11348595.
In agreement with these results, the proposed generation of double strand breaks in pol3-exo(-) rad27 mutants was suppressed by the overexpression of DNA2. Paper-9868442.
Purified Mph1 stimulated the endonuclease activities of both Fen1 and Dna2, which act faithfully in the processing of Okazaki fragments. Paper-13714835.
To determine why the disengagement mechanism evolved, we measured FEN1 dissociation of Dna2 on short RNA and DNA flaps, which occur during flap processing. Paper-13677576.
Here we show that the helicase domain is required in vivo and that a 3' to 5' DNA helicase activity specific for forked substrates is intrinsic to the Dna2p. Paper-416672.
In addition, we identified physical and functional interactions between these proteins and found that RPA binds Dna2 predominantly through its large subunit, Rpa1. Paper-9718988.
Replication protein A (RPA)-bound flaps inhibit cleavage by FEN1 but stimulate Dna2, requiring that Dna2 cleaves prior to FEN1. Paper-13677576.
Temperature-sensitive (Ts) dna2 mutants arrest irreversibly at G2/M in a RAD9- and MEC1-dependent manner, suggesting that Dna2p has a role in S phase. Paper-8260933.
RFA1, the gene encoding the large subunit of RPA, displayed allele-specific interactions with DNA2 that included synthetic lethality and intergenic complementation. Paper-9718988.
In addition, the life span of dna2 mutants is extended by expression of an additional copy of SIR2 or by deletion of FOB1, which also increase wild-type life span. Paper-9184851.
Multicopy expression of MPH1 also suppressed the temperature-sensitive growth defects in cells expressing dna2Delta405N, which lacks the N-terminal 405 amino acids of Dna2. Paper-13714835.
As with its role in cell cycle progression, both the N-terminal and C-terminal regions, as well as the kinase domain of Tor1p, are required for rescue of dna2 mutants. Paper-8260933.
In this study, we identified MGS1 as a multicopy suppressor of the temperature-sensitive dna2Delta405N mutation, a DNA2 allele lacking the N-terminal 405 amino acid residues. Paper-10977272.
In the other, the single-stranded binding protein, replication protein A (RPA), coats the flap, inhibits FEN1, but stimulates cleavage by the Dna2p helicase/nuclease. Paper-10238035.
Arrest of dna2 mutants was RAD9 dependent, but deleting this checkpoint resulted in either no effect or suppression of defects, including the synthetic lethality with ctf4. Paper-1823220.
Using this novel system we demonstrate that semi-conservative replication, but not polymerase activity per se, requires the activity of the DNA helicase encoded by DNA2. Paper-1557297.
Dna2 mutants reveal interactions with Dna polymerase alpha and Ctf4, a Pol alpha accessory factor, and show that full Dna2 helicase activity is not essential for growth. Paper-1823220.
In summary, RPA, Dna2, and FEN1 have evolved highly coordinated binding properties enabling one protein to succeed the next for proper and efficient Okazaki flap processing. Paper-13677576.
Additional results showed that flap substrate recognition and tracking by FEN1, as occur during fragment processing, are required for effective displacement of the flap-bound Dna2. Paper-13677576.
Our biochemical and genetic data indicate that the in vivo suppression of Dna2 defects observed with both dna2K1080E and dna2Delta405N mutants occur via stimulation of Fen1 activity. Paper-13714835.
Homology searches revealed protein sequence homology between HEL1 and two previously identified and biochemically characterized yeast helicases, encoded by the DNA2 and UPF1 genes. Paper-1302600.
Additional mutations known or expected to destroy the ATPase and helicase activities of Dna2 were constructed and found to support growth on some media but to cause alkylation sensitivity. Paper-1823220.
To investigate which of these activities is important for the cellular functions of Dna2, we have identified separation of function mutations that selectively inactivate the helicase or nuclease. Paper-8461992.
Deletion of SGS1 or DNA2 reduces resection and DSB repair by single-strand annealing between distant repeats while the remaining long-range resection activity depends on the exonuclease Exo1. Paper-12950376.
Telomeric localization of Dna2p required Sir3p, since the amount of Dna2p found at telomeres by two different assays, one-hybrid and ChIP, is severely reduced in strains lacking Sir3p. Paper-9209910.
Saccharomyces cerevisiae Dna2 protein is required for DNA replication and repair and is associated with multiple biochemical activities: DNA-dependent ATPase, DNA helicase, and DNA nuclease. Paper-8461992.
The presence of high Dna2p activity, under reaction conditions favoring helicase activity, substantially stimulated FEN1 cleavage of tailed-foldback flaps and also 30-nucleotide unstructured flaps. Paper-10238035.
A point mutation at the conserved ATP-binding site of Dna2 inactivated concurrently ssDNA-dependent ATPase, ATP-dependent nuclease, and helicase activities, indicating that they all reside in Dna2 itself. Paper-1597805.
Our data thus provide in vivo evidence for the types of DNA lesions predicted to occur when lagging-strand synthesis is deficient and suggest that Dna2p and Rad27p collaborate in the processing of Okazaki fragments. Paper-1867060.
For example, the DNA-binding domain of the yeast GAL4 protein interacts very poorly with nucleosome cores compared with naked DNA2 (and see below), and binding of other activators is even more strongly inhibited. Paper-127614.
Moreover, the N-terminal domains of Dna2 and Rpa1 appear to be important for a functional interaction because the N-terminal domain of RPA1 was required to maximally stimulate Dna2 endonuclease activity. Paper-9718988.
Overproduction of Dna2 suppresses the rad27/ rth1 delta temperature-sensitive growth defect. dna2-1 rad27/ rth1 delta double mutants are inviable, indicating that the mutations are synthetically lethal. Paper-958038.
Analysis of the Cd(2+)-sensitivity of RAD27 (rad27-G67S) and DNA2 ( dna2-1) separation of function alleles revealed that their activities necessary for Okazaki fragment processing are essential in conditions of cadmium exposure. Paper-12881956.
In characterizing dna2 mutants, we have found that Dna2p also participates in DNA repair or in damage avoidance mechanisms. dna2 mutants are sensitive to X rays, although they are less sensitive than rad52 mutants. Paper-8463768.
The X-ray sensitivity of dna2 mutants is suppressed by overexpression of a 5' to 3' exonuclease, the yeast FEN-1 structure-specific nuclease, encoded by the RAD27 gene, which also suppresses the growth defect of dna2-ts mutants. Paper-8463768.
Overexpression of Dna2p in wild-type cells also yielded single-stranded DNA regions on telomeric DNA and caused a cell growth arrest phenotype virtually identical to that seen for rad27 cells grown at the restrictive temperature. Paper-1867060.
All of the phenotypes of old wild-type cells, for example, extended cell cycle time, age-related transcriptional silencing defects, and nucleolar reorganization, occur after fewer generations in dna2 mutants than in the wild type. Paper-9184851.
To gain further insights into the biological functions of Dna2, previously known as a cellular replicative helicase in Saccharomyces cerevisiae, we examined biochemical properties of the recombinant Dna2 protein purified to homogeneity. Paper-1597805.
Using a nuclease-defective Dna2 mutant, we reconstituted the sequential dissociation reactions in the proposed RPA/ Dna2/ FEN1 pathway showing that, even without cutting, Dna2 enables FEN1 to cleave RPA-coated flaps. Paper-13677576.
We further show that deletion of DNA2 suppresses the long-telomere phenotype and the high rate of formation of gross chromosomal rearrangements in pif1Delta mutants, suggesting a role for Dna2 in telomere elongation in the absence of Pif1. Paper-11348595.
One proposed pathway for flap removal involves pol delta displacement of long flaps, coating of those flaps by replication protein A ( RPA), and sequential cleavage of the flap by Dna2 nuclease followed by flap endonuclease 1 ( FEN1). Paper-12209380.
In this work, we present evidence that Saccharomyces cerevisiae Pif1 helicase plays a wider role in DNA replication than previously appreciated and that it likely functions in conjunction with Dna2 helicase/nuclease as a component of the OFP machinery. Paper-11348595.
The DNA2 gene is essential and encodes a 172-kDa protein with DNA helicase motifs in its C-terminal half and an N-terminal half with no similarity to any previously described protein (Budd, M. E., and Campbell, J. L. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 7642-7646). Paper-416672.
The search for NLSs that overlap the consensus CDK1 phosphorylation site by using cNLS Mapper identified all previously reported and 5 previously uncharacterized yeast proteins ( Yen1, Psy4, Pds1, Msa1, and Dna2) displaying CDK1- and cell cycle-regulated nuclear transport. Paper-13856260.
Both the helicase and nuclease activities of Dna2Pho were inhibited by substrates with RNA segments at the 5'-end of flap DNA, whereas the nuclease activity of Dna2 from S. cerevisiae was reported to be stimulated by RNA segments in the 5'-tail (Bae, S.-H., and Seo, Y. S. (2000) J. Biol. Chem. 38022-38031). Paper-9695890.
Mutations in the gene for the conserved, essential nuclease-helicase Dna2 from the yeast Saccharomyces cerevisiae were found to interact genetically with POL1 and CTF4, which encode a DNA Polymerase alpha subunit and an associated protein, suggesting that Dna2 acts in a process that involves Pol alpha. Paper-1823220.
We have previously shown that replication- protein A (RPA), the heterotrimeric single-stranded DNA binding protein of eukaryotes, plays a role in Okazaki fragment processing by acting as a molecular switch between the two endonucleases, Dna2 and Fen1, to ensure the complete removal of primer RNAs in Saccharomyces cerevisiae. Paper-9718988.
Additional genetic interactions were observed between pob3 mutations and the genes encoding several DNA replication factors, including POL1, CTF4, DNA2, and CHL12. pob3 alleles caused sensitivity to the ribonucleotide reductase inhibitor hydroxyurea, indicating a defect in a process requiring rapid dNTP synthesis. Paper-8448411.
We report here that the absence of Rad27 and Dna2 functions but not RNase H(35) or Exo1, which play an essential role in the processing of Okazaki fragments during replication, destabilize the human minisatellite CEB1 in mitotically growing Saccharomyces cerevisiae cells, up to 14% per generation in rad27Delta cells. Paper-9486766.
These synonyms are used for gene DNA2 (Dna2p): YHR164C, DNA replication ATP-dependent helicase DNA2.
These accession numbers are used for gene DNA2: AAB68010 (NCBI_GENBANK__AC).
DNA2 is a homologue of KLLA0A05324g (hypothetical protein) from Kluyveromyces lactis NRRL Y-1140.
DNA2 is a homologue of AGOS_AEL218W (AEL218Wp) from Ashbya gossypii ATCC 10895.
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