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(Received for publication, December 14, 1994 ) From the
A 5`- to 3`-exonuclease of about 45 kDa has been purified
from various mammalian sources and shown to be required for the
completion of lagging strand synthesis in reconstituted DNA replication
systems. RTH1 encodes the yeast Saccharomyces cerevisiae counterpart of the mammalian enzyme. To determine the in vivo biological role of RTH1-encoded 5`- to 3`-exonuclease, we
have examined the effects of an rth1 In Escherichia coli, the 5`- to 3`-exonuclease activity
of DNA polymerase I is involved in the removal of RNA primers attached
to the 5`-end of newly replicated DNA. The E. coli polA ex1 mutant is defective in 5`- to 3`-exonuclease activity and is
temperature-sensitive for growth. The joining of Okazaki fragments is
retarded in the polA ex1 mutant, and as a consequence, it
exhibits elevated levels of genetic recombination (1, 2) . In eukaryotes, a 5`- to 3`-exonuclease
with a molecular size of 45 kDa has been purified from HeLa cells,
mouse, and calf thymus(3, 4, 5) . Ishimi et al.(4) reconstituted the replication of simian
virus 40 origin containing DNA using SV40 large T antigen and other
purified components (single-stranded DNA binding protein RPA, DNA
polymerase The mouse and human 5`- to 3`-exonuclease
genes have been cloned, and they encode highly homologous proteins of
378 and 380 amino acids, respectively(8, 9) . Both
proteins share a high degree of homology with the Saccharomyces
cerevisiae protein of 382 amino acids encoded by the YKL510 open
reading frame(8, 9) , and the S. cerevisiae protein also has a 5`- to 3`-exonuclease activity(8) . All
these exonucleases share homology with the S. cerevisiae and
human nucleotide excision repair proteins RAD2 and XPG, respectively.
Both the RAD2 and XPG proteins contain DNA endonuclease and 5`- to
3`-exonuclease activities(10, 11, 12) . RAD2
and XPG, however, are much larger proteins containing 1031 and 1186
residues, respectively, and the homology between RAD2/XPG and the above
noted mammalian and yeast 5`- to 3`-exonucleases occurs in three
regions(8, 13, 14) . Because of the homology
of the S. cerevisiae YKL510 open reading frame encoded protein
with RAD2, we have named the gene contained within the YKL510 DNA
fragment RTH1 (RAD two homolog). The strict
dependence of reconstituted mammalian DNA replication systems on the
5`- to 3`-exonuclease suggested that RTH1 may be an essential
gene. To define the biological role of RTH1, we have examined
the effects of a null mutation in RTH1 on viability, cell
cycle morphology, and spontaneous mitotic recombination. We find that
the rth1
For
Three 10-ml cultures were
grown to a density of 10
For examining terminal morphology
at 37 °C, following ethanol fixation, cells were rehydrated in 40
mM KPO
Figure 1:
UV survival
of rth1
The rth1
Figure 2:
Lethality of rth1
In vitro reconstitution of the DNA replication machinery from
different mammalian sources has indicated the requirement of a 5`- to
3`-exonuclease activity in the completion of lagging strand DNA
synthesis. RTH1 encodes the S. cerevisiae counterpart
of the mammalian 5`- to 3`-exonuclease. In this study, we have
determined the in vivo biological role of RTH1 by
examining the effects of the rth1 As expected for a mutant
defective in DNA replication, the rth1 We find no evidence for the involvement of RTH1 in the
repair of UV damage. The rth1
Volume 270,
Number 9,
Issue of March 3, 1995 pp. 4193-4196
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
mutation on various
cellular processes. rth1 mutants grow poorly at 30
°C, and a cessation in growth occurs upon transfer of the mutant to
37 °C. At the restrictive temperature, the rth1 mutant exhibits a terminal cell cycle morphology similar to that
of mutants defective in DNA replication, and levels of spontaneous
mitotic recombination are elevated in the rth1 mutant
even at the permissive temperature. The rth1 mutation
does not affect UV or 
-ray sensitivity but enhances sensitivity to
the alkylating agent methyl methanesulfonate. The role of RTH1 in DNA replication and in repair of alkylation damage is
discussed.
-primase complex, topoisomerase II, ribonuclease H, 5`-
to 3`-exonuclease, and DNA ligase) isolated from HeLa cells. The 5`- to
3`-exonuclease was essential for the production of replicating form I
DNA. A 5`- to 3`-exonuclease from mouse cells has also been shown to be
required in a pol-primase-dependent replication that used
single-stranded circular DNA as a template(5) . Both of these
studies indicated that the removal of RNA primers required the combined
action of RNase H and 5`- to 3`-exonuclease. In another
study(6) , the 5`- to 3`-exonuclease has been shown to
functionally interact with DNA polymerase , 
, or 
in the
completion of lagging strand DNA synthesis. Using purified proteins
from calf thymus, Turchi et al.(7) have shown that
RNase H1 cleaves the primer RNA one nucleotide 5` of the RNA-DNA
junction, and the remaining monoribonucleotide is removed by the 5`- to
3`-exonuclease activity. mutation is conditionally lethal, and at the
restrictive temperature, mutant cells exhibit a cell cycle morphology
characteristic of mutants defective in DNA replication.
Construction of a Null Mutation of the RTH1
Gene
The plasmid pR2.10, a derivative of pUC19, was used to make
a genomic deletion of the RTH1 gene. pR2.10 contains the RTH1 gene in which nucleotides +58 to +755 of the RTH1 1146 nucleotide open reading frame (15) have been
replaced with the yeast URA3 gene flanked by Salmonella
typhimurium HisG sequences(16) . An rth1 mutation was made by the gene replacement technique (17) by cutting pR2.10 with EcoRI and SalI
and transforming yeast cells to Ura
. Genomic deletion
mutations of RTH1 and various RAD genes were
constructed in S. cerevisiae strains EMY6 (MAT ade5
his7 leu2 lys1 met14 trp1 ura3) and LP3041-6D (MATaleu2-3 leu2-112 trp1 ura3-52). Sensitivity to UV and
Quantification of survival after UV irradiation of
yeast strains was carried out as described(18) . Cultures were
grown from single colonies to stationary phase in the appropriate
selection media. Cells were sonicated for 1-3 min in a Branson
water bath sonicator, diluted, and plated on the appropriate medium.
Plates were exposed to UV irradiation, incubated at 25, 30, or 34
°C for 3-5 days in the dark to avoid photoreactivation, and
the colonies were counted.
Irradiation-ray irradiation, RTH1 and rth1 strains were suspended in U-wells and
transferred to yeast extract-peptone-dextrose (YPD) (
)plates. Following irradiation with a cobalt-60 source at a
dose rate of 9 kilorads/min, plates were incubated at either 25, 30, or
34 °C for 3-4 days and examined at regular intervals. Sensitivity to the DNA Alkylating Agent Methyl
Methanesulfonate (MMS)
RTH1 and rth1 strains were transferred to YPD plates containing either no MMS or
MMS concentrations ranging from 0.02 to 0.035%. Plates were incubated
at 25, 30, or 34 °C for 4-5 days and examined. Rates of Mitotic Recombination
The his33`, his35` duplication was constructed in wild
type strains LP3041-6D, EMY6, and their rth1 derivatives by introducing the plasmid pRS6, which contains an
internal fragment of the HIS3 gene, the LEU2 gene,
and pBR322 sequences. Integration of pRS6 at the genomic HIS3 site results in two copies of the his3 gene, one with a
terminal deletion at the 3`-end and the other with a terminal deletion
at the 5`-end. The two his3 alleles are separated by LEU2 and pBR322 sequences(19) .
cells/ml at either 25, 30, or 34
°C in synthetic liquid medium lacking leucine. As nearly all of the HIS3 recombinants show a simultaneous loss of
the LEU2 gene and pBR322 sequences, HIS3 recombinants do not divide in medium lacking leucine. Therefore,
the frequency of HIS3 recombinants measures
the actual recombination rate. Log phase cultures were washed,
resuspended in 1 ml of sterile water, diluted, and plated onto
synthetic complete media to determine viability and onto synthetic
complete media lacking histidine to determine the frequency of HIS3 recombinants. Plates were incubated for
3-4 days at 25, 30, or 34 °C, and the frequency of HIS3 recombinants was determined.
Recombination rates were calculated by the method of median of Lea and
Coulson (as described previously in (19) ). Growth Rates and Cell Morphology
Doubling
times were determined from growth curves obtained for the various yeast
strains at 30 °C in liquid YPD medium. Cultures were diluted to A
of about 0.05, and the densities followed to A of 4-5. For growth at 37 °C,
cultures were divided with half of the culture left at 30 °C and
the other half shifted to 37 °C and A determined at various times.
, pH 6.5 buffer containing 0.5 M MgCl
and 1.2 M sorbitol, and mounted on
slides coated with 0.1% polylysine (M
400,000).
For visualization of nuclear morphology, a drop of mounting medium
containing 4`,6`-diamidino-2-phenylindole was applied to the slide, and
cells were photographed with a Leitz Laborlux D fluorescence microscope
equipped with an Olympus PM-10AD photomicrographic system.
Sensitivity of the rth1
To test if RTH1 has a role in DNA repair, we
examined the sensitivity of the rth1 Mutation to DNA Damaging
Agents mutant to
ultraviolet irradiation, -irradiation, and the alkylating agent
MMS. The rth1 mutation was constructed in two different
yeast strains and their sensitivity to DNA damaging agents examined at
three different temperatures, 25, 30, and 34 °C. As shown in Fig. 1, A and B, the rth1 mutation has no effect on UV sensitivity. The UV sensitivity of
the rth1 mutation, in combination with the rad2 mutation defective in nucleotide excision repair, is the same as
that of the rad2 single mutation, suggesting that RTH1 has no significant role in the RAD6 postreplicative bypass or the RAD52 recombinational
repair pathways, which, in addition to nucleotide excision repair,
constitute the three pathways for the repair of UV damage in
yeast(20) . We also found no effect of the rth1 mutation on -ray sensitivity (data not shown). Sensitivity to
MMS was, however, enhanced by the rth1 mutation (Fig. 1C).
, rad2, and rad2 rth1 strains. 
, wild type strain; 
, rth1;
, rad2; 
, rad2 rth1. A,
wild type strain LP3041-6D and its rth1, rad2, and rth1 rad2 derivative mutant
strains were grown at 30 °C, and cells were plated and
UV-irradiated, followed by incubation of plates at 30 °C in the
dark. Similar results were obtained at 25 or 34 °C. B,
wild type strain EMY6 and its mutant derivatives were grown at 34
°C and following UV irradiation plates were incubated at 34 °C
in the dark. Similar results were observed at 25 or 30 °C. C, MMS sensitivity of rth1 mutant. The wild type (WT) strain EMY6 and its rth1 derivative were
grown on YPD + 0.03% MMS (+ MMS) or on YPD medium lacking MMS
(-MMS) at 30 °C. Similar results were observed for
LP3041-6D and its rth1 derivative.
Conditional Lethality of the rth1
When grown at 30 °C, the rth1
Mutation mutation affects growth rate considerably, increasing the cell
doubling time to twice to that in the wild type strain (Table 1).
We also determined the growth rate of the rth1 mutation
in combination with mutations in genes representing the three epistasis
groups for the repair of UV damage in S. cerevisiae (Table 1). The growth rates of the rth1 rad2 and rth1 rad6 double mutants were about the
same as that of the rth1 single mutant; the doubling time
of the rth1 rad52 mutant was, however, about twice
that of the rth1 or the rad52 single
mutant, indicating that the RAD52 recombinational pathway
plays an important role in repairing the DNA lesions that accumulate in
the rth1 mutant.
mutant stops
growth upon transfer to 37 °C (Fig. 2A). Microscopic
examination revealed that rth1 mutant cells stop division as
two large cells consisting of the mother and the daughter cell, with a
block in nuclear division. In some cases, the nucleus has migrated to
the neck between the two cells, whereas in other cases, cells exhibit
an elongated nucleus stretching between the mother and daughter cell (Fig. 2B). The cell cycle arrest phenotype of the rth1 mutant resembles that of the various S.
cerevisiae mutants known to be defective in DNA replication. For
example, cdc2 mutants with a defect in DNA polymerase
stop at the restrictive temperature with the nucleus that has migrated
to the neck of the mother cell but has not elongated, whereas the DNA
ligase defective mutant cdc9 stops cell division with an
elongated nucleus extending between the two cells(21) .
mutant at
37 °C. A, rth1 mutation inhibits growth at 37 °C.
Log phase cultures of LP3041-6D and its rth1 derivative were diluted to approximately A = 0.05, and the cultures were then split. One half of the
culture was incubated at 30 °C and the other half at 37 °C.
, RTH1 (30 °C); , RTH1 (37 °C);
, rth1 (30 °C); , rth1 (37 °C). B, cell cycle morphology of rth1 strain. The terminal morphology of rth1 cells was
examined by 4`,6`-diamidino-2-phenylindole
staining.
Elevated Rates of Spontaneous Mitotic Recombination
in the rth1
In E. coli, mutations in the
5`- to 3`-exonuclease activity of DNA polymerase I and in DNA ligase
confer a hyper recombination phenotype because of the retarded joining
of Okazaki fragments(1, 2, 22) . In S.
cerevisiae, mutations in DNA replication genes encoding DNA
ligase(23) , DNA polymerases, and other enzymes (24) also result in elevated rates of spontaneous mitotic
recombination. The requirement of 5`- to 3`-exonuclease encoded by the
mammalian counterparts of RTH1 in the completion of lagging
strand synthesis in reconstituted DNA replication systems suggested
that retarded joining of Okazaki fragments in rth1 Mutant mutant
cells should result in an increased level of spontaneous mitotic
recombination. To verify this, we examined the effect of the rth1 mutation on spontaneous mitotic intrachromosomal
recombination between two his3 genes (his33`,
his35`) in two different RAD strains and in their rth1 derivatives. As shown in Table 2, the rate of mitotic recombination is elevated
15-30-fold in the rth1 mutant.
mutation on viability,
mitotic recombination, and DNA repair. We find that RTH1 is
not an essential gene. However, growth rate is slowed very considerably
in the rth1 mutant at the permissive temperature, and the rth1 mutation is inviable at the restrictive temperature
of 37 °C. These results suggest the presence of an alternate 5`- to
3`-exonuclease activity that, at the permissive temperature, can
substitute for the activity missing in the rth1 mutant;
however, at the elevated temperature, the other 5`- to 3`-exonuclease
activity is unable to support DNA replication. Because of the homology
of RTH1-encoded protein with the S. cerevisiae RAD2
protein and the fact that RAD2 also possesses a 5`- to 3`-exonuclease
activity(10) , we determined the effect of the rad2 mutation on viability in combination with the rth1 mutation. However, the rad2 mutation has no effect
on viability or growth rate of the rth1 mutation,
indicating that RAD2 does not fulfill the role of the alternate 5`- to
3`-exonuclease in DNA replication. mutant stops
division at the restrictive temperature as two large cells with a
defect in nuclear division. At the permissive temperature, the rth1 mutation results in a reduction in growth rate, and
a further decline in growth rate occurs in the rth1 rad52 double mutant. A slowdown in the removal of RNA primers in the rth1 mutant would retard the joining of nascent DNA
fragments; subsequent channeling of these discontinuities into the RAD52 recombinational repair pathway would result in elevated
levels of spontaneous mitotic recombination observed in the rth1 mutant. Elimination of recombinational repair by the rad52 mutation would leave the DNA lesions in the rth1 mutant unrepaired, resulting in a further reduction
in growth rate of rth1 rad52 mutant strain over that
of the rth1 and rad52 single mutants. We
have shown previously that DNA ligase-deficient mutations are lethal in
combination with mutations in the RAD52 gene(23) . mutation, however, confers
sensitivity to MMS, suggesting a role for the RTH1 5`- to
3`-exonuclease in the repair of alkylation damage. Following removal of
the alkylated base by a DNA glycosylase and subsequent cleavage of the
phosphodiester bond at the 5`-side of the apurinic/apyrimidinic (AP)
residue by a class II AP endonuclease, the 5`- to 3`-exonuclease could
effect the release of the AP residue, resulting in a 1-nucleotide gap,
which could then be filled in by repair synthesis.
)
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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S. Tom, L. A. Henricksen, and R. A. Bambara Mechanism Whereby Proliferating Cell Nuclear Antigen Stimulates Flap Endonuclease 1 J. Biol. Chem., March 31, 2000; 275(14): 10498 - 10505. [Abstract] [Full Text] [PDF]
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Q. Liu, W.-c. Choe, and J. L. Campbell Identification of the Xenopus laevis Homolog of Saccharomyces cerevisiae DNA2 and Its Role in DNA Replication J. Biol. Chem., January 21, 2000; 275(3): 1615 - 1624. [Abstract] [Full Text] [PDF]
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M. U. Fikus, P. A. Mieczkowski, P. Koprowski, J. Rytka, E. ledziewska-Gójska, and Z. Ciela The Product of the DNA Damage-Inducible Gene of Saccharomyces cerevisiae, DIN7, Specifically Functions in Mitochondria Genetics, January 1, 2000; 154(1): 73 - 81. [Abstract] [Full Text]
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B.-I. Lee and D. M. Wilson III The RAD2 Domain of Human Exonuclease 1 Exhibits 5' to 3' Exonuclease and Flap Structure-specific Endonuclease Activities J. Biol. Chem., December 31, 1999; 274(53): 37763 - 37769. [Abstract] [Full Text] [PDF]
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J. Qiu, Y. Qian, P. Frank, U. Wintersberger, and B. Shen Saccharomyces cerevisiae RNase H(35) Functions in RNA Primer Removal during Lagging-Strand DNA Synthesis, Most Efficiently in Cooperation with Rad27 Nuclease Mol. Cell. Biol., December 1, 1999; 19(12): 8361 - 8371. [Abstract] [Full Text] [PDF]
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R. Gary, M. S. Park, J. P. Nolan, H. L. Cornelius, O. G. Kozyreva, H. T. Tran, K. S. Lobachev, M. A. Resnick, and D. A. Gordenin A Novel Role in DNA Metabolism for the Binding of Fen1/Rad27 to PCNA and Implications for Genetic Risk Mol. Cell. Biol., August 1, 1999; 19(8): 5373 - 5382. [Abstract] [Full Text] [PDF]
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P. J. White, R. H. Borts, and M. C. Hirst Stability of the Human Fragile X (CGG)n Triplet Repeat Array in Saccharomyces cerevisiae Deficient in Aspects of DNA Metabolism Mol. Cell. Biol., August 1, 1999; 19(8): 5675 - 5684. [Abstract] [Full Text] [PDF]
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E. Matsui, S. Kawasaki, H. Ishida, K. Ishikawa, Y. Kosugi, H. Kikuchi, Y. Kawarabayashi, and I. Matsui Thermostable Flap Endonuclease from the Archaeon, Pyrococcus horikoshii, Cleaves the Replication Fork-like Structure Endo/Exonucleolytically J. Biol. Chem., June 25, 1999; 274(26): 18297 - 18309. [Abstract] [Full Text] [PDF]
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J. Qiu, Y. Qian, V. Chen, M.-X. Guan, and B. Shen Human Exonuclease 1 Functionally Complements Its Yeast Homologues in DNA Recombination, RNA Primer Removal, and Mutation Avoidance J. Biol. Chem., June 18, 1999; 274(25): 17893 - 17900. [Abstract] [Full Text] [PDF]
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J. Parenteau and R. J. Wellinger Accumulation of Single-Stranded DNA and Destabilization of Telomeric Repeats in Yeast Mutant Strains Carrying a Deletion of RAD27 Mol. Cell. Biol., June 1, 1999; 19(6): 4143 - 4152. [Abstract] [Full Text] [PDF]
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 F. Paques and J. E. Haber Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae Microbiol. Mol. Biol. Rev., June 1, 1999; 63(2): 349 - 404. [Abstract] [Full Text] [PDF]
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J. A. Rumbaugh, L. A. Henricksen, M. S. DeMott, and R. A. Bambara Cleavage of Substrates with Mismatched Nucleotides by Flap Endonuclease-1. IMPLICATIONS FOR MAMMALIAN OKAZAKI FRAGMENT PROCESSING J. Biol. Chem., May 21, 1999; 274(21): 14602 - 14608. [Abstract] [Full Text] [PDF]
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T. Formosa and T. Nittis 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 Genetics, April 1, 1999; 151(4): 1459 - 1470. [Abstract] [Full Text]
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R. Gary, K. Kim, H. L. Cornelius, M. S. Park, and Y. Matsumoto Proliferating Cell Nuclear Antigen Facilitates Excision in Long-patch Base Excision Repair J. Biol. Chem., February 12, 1999; 274(7): 4354 - 4363. [Abstract] [Full Text] [PDF]
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C.-Y. Kim, B. Shen, M. S. Park, and G. A. Olah Structural Changes Measured by X-ray Scattering from Human Flap Endonuclease-1 Complexed with Mg2+ and Flap DNA Substrate J. Biol. Chem., January 15, 1999; 274(3): 1233 - 1239. [Abstract] [Full Text] [PDF]
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G. Frank, J. Qiu, M. Somsouk, Y. Weng, L. Somsouk, J. P. Nolan, and B. Shen Partial Functional Deficiency of E160D Flap Endonuclease-1 Mutant in Vitro and in Vivo Is Due to Defective Cleavage of DNA Substrates J. Biol. Chem., December 4, 1998; 273(49): 33064 - 33072. [Abstract] [Full Text] [PDF]
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J. A. Rumbaugh, G. M. Fuentes, and R. A. Bambara Processing of an HIV Replication Intermediate by the Human DNA Replication Enzyme FEN1 J. Biol. Chem., October 30, 1998; 273(44): 28740 - 28745. [Abstract] [Full Text] [PDF]
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D. J. Hosfield, G. Frank, Y. Weng, J. A. Tainer, and B. Shen Newly Discovered Archaebacterial Flap Endonucleases Show a Structure-Specific Mechanism for DNA Substrate Binding and Catalysis Resembling Human Flap Endonuclease-1 J. Biol. Chem., October 16, 1998; 273(42): 27154 - 27161. [Abstract] [Full Text] [PDF]
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J. J. Miret, L. Pessoa-Brandao, and R. S. Lahue Orientation-dependent and sequence-specific expansions of CTG/CAG trinucleotide repeats in Saccharomyces cerevisiae PNAS, October 13, 1998; 95(21): 12438 - 12443. [Abstract] [Full Text] [PDF]
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R. J. Kokoska, L. Stefanovic, H. T. Tran, M. A. Resnick, D. A. Gordenin, and T. D. Petes Destabilization of Yeast Micro- and Minisatellite DNA Sequences by Mutations Affecting a Nuclease Involved in Okazaki Fragment Processing (rad27) and DNA Polymerase delta (pol3-t) Mol. Cell. Biol., May 1, 1998; 18(5): 2779 - 2788. [Abstract] [Full Text]
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K. Kim, S. Biade, and Y. Matsumoto Involvement of Flap Endonuclease 1 in Base Excision DNA Repair J. Biol. Chem., April 10, 1998; 273(15): 8842 - 8848. [Abstract] [Full Text] [PDF]
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 C. H. Freudenreich, S. M. Kantrow, and V. A. Zakian Expansion and Length-Dependent Fragility of CTG Repeats in Yeast Science, February 6, 1998; 279(5352): 853 - 856. [Abstract] [Full Text]
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D. F. Fiorentino and G. R. Crabtree Characterization of Saccharomyces cerevisiae dna2 Mutants Suggests a Role for the Helicase Late in S Phase Mol. Biol. Cell, December 1, 1997; 8(12): 2519 - 2537. [Abstract] [Full Text]
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M. Bhagwat, L. J. Hobbs, and N. G. Nossal The 5'-Exonuclease Activity of Bacteriophage T4 RNase H Is Stimulated by the T4 Gene 32 Single-stranded DNA-binding Protein, but Its Flap Endonuclease Is Inhibited J. Biol. Chem., November 7, 1997; 272(45): 28523 - 28530. [Abstract] [Full Text] [PDF]
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J. A. Rumbaugh, R. S. Murante, S. Shi, and R. A. Bambara Creation and Removal of Embedded Ribonucleotides in Chromosomal DNA during Mammalian Okazaki Fragment Processing J. Biol. Chem., September 5, 1997; 272(36): 22591 - 22599. [Abstract] [Full Text] [PDF]
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R. A. Bambara, R. S. Murante, and L. A. Henricksen Enzymes and Reactions at the Eukaryotic DNA Replication Fork J. Biol. Chem., February 21, 1997; 272(8): 4647 - 4650. [Full Text] [PDF]
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M. Yao and Y. W. Kow Cleavage of Insertion/Deletion Mismatches, Flap and Pseudo-Y DNA Structures by Deoxyinosine 3'-Endonuclease from Escherichia coli J. Biol. Chem., November 29, 1996; 271(48): 30672 - 30676. [Abstract] [Full Text] [PDF]
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C. J. Barnes, A. F. Wahl, B. Shen, M. S. Park, and R. A. Bambara Mechanism of Tracking and Cleavage of Adduct-damaged DNA Substrates by the Mammalian 5'- to 3'-Exonuclease/Endonuclease RAD2 Homologue 1or Flap Endonuclease 1 J. Biol. Chem., November 22, 1996; 271(47): 29624 - 29631. [Abstract] [Full Text] [PDF]
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M. S. DeMott, B. Shen, M. S. Park, R. A. Bambara, and S. Zigman Human RAD2 Homolog 15'- to 3'-Exo/Endonuclease Can Efficiently Excise a Displaced DNA Fragment Containing a 5'-Terminal Abasic Lesion by Endonuclease Activity J. Biol. Chem., November 22, 1996; 271(47): 30068 - 30076. [Abstract] [Full Text] [PDF]
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R. S. Murante, J. A. Rumbaugh, C. J. Barnes, J. R. Norton, and R. A. Bambara Calf RTH-1 Nuclease Can Remove the Initiator RNAs of Okazaki Fragments by Endonuclease Activity J. Biol. Chem., October 18, 1996; 271(42): 25888 - 25897. [Abstract] [Full Text] [PDF]
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 R Kolodner Biochemistry and genetics of eukaryotic mismatch repair. Genes & Dev., June 15, 1996; 10(12): 1433 - 1442. [PDF]
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B. Shen, J. P. Nolan, L. A. Sklar, and M. S. Park Essential Amino Acids for Substrate Binding and Catalysis of Human Flap Endonuclease 1 J. Biol. Chem., April 19, 1996; 271(16): 9173 - 9176. [Abstract] [Full Text] [PDF]
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Y. Habraken, P. Sung, L. Prakash, and S. Prakash Structure-specific Nuclease Activity in Yeast Nucleotide Excision Repair Protein Rad2 J. Biol. Chem., December 15, 1995; 270(50): 30194 - 30198. [Abstract] [Full Text] [PDF]
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M. E. Budd, W.-C. Choe, and J. L. Campbell DNA2 Encodes a DNA Helicase Essential for Replication of Eukaryotic Chromosomes J. Biol. Chem., November 10, 1995; 270(45): 26766 - 26769. [Abstract] [Full Text] [PDF]
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X. Li, J. Li, J. Harrington, M. R. Lieber, and P. M. J. Burgers Lagging Strand DNA Synthesis at the Eukaryotic Replication Fork Involves Binding and Stimulation of FEN-1 by Proliferating Cell Nuclear Antigen J. Biol. Chem., September 22, 1995; 270(38): 22109 - 22112. [Abstract] [Full Text] [PDF]
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R. Johnson, G. Kovvali, L Prakash, and S Prakash Requirement of the yeast RTH1 5' to 3' exonuclease for the stability of simple repetitive DNA Science, July 14, 1995; 269(5221): 238 - 240. [Abstract] [PDF]
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S.-H. Bae and Y.-S. Seo Characterization of the Enzymatic Properties of the Yeast Dna2 Helicase/Endonuclease Suggests a New Model for Okazaki Fragment Processing J. Biol. Chem., November 22, 2000; 275(48): 38022 - 38031. [Abstract] [Full Text] [PDF]
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M. Stucki, Z. O. Jonsson, and U. Hubscher In Eukaryotic Flap Endonuclease 1, the C Terminus Is Essential for Substrate Binding J. Biol. Chem., March 9, 2001; 276(11): 7843 - 7849. [Abstract] [Full Text] [PDF]
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M. Bhagwat and N. G. Nossal Bacteriophage T4 RNase H Removes Both RNA Primers and Adjacent DNA from the 5' End of Lagging Strand Fragments J. Biol. Chem., July 20, 2001; 276(30): 28516 - 28524. [Abstract] [Full Text] [PDF]
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L. A. Henricksen, S. Tom, Y. Liu, and R. A. Bambara Inhibition of Flap Endonuclease 1 by Flap Secondary Structure and Relevance to Repeat Sequence Expansion J. Biol. Chem., May 26, 2000; 275(22): 16420 - 16427. [Abstract] [Full Text] [PDF]
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