ライフサイエンスコーパス: FEN-1

1 FEN-1 also appears to be important in mismatch repair.

2 FEN-1 binds the unpaired 3' DNA end (3' flap), opens and

3 FEN-1 consists of a nuclease core domain and a C-termina

4 FEN-1 has specific endonuclease activity for repairing n

5 FEN-1 homologues from humans, Saccharomyces cerevisiae,

6 FEN-1 is essential for lagging strand DNA synthesis in O

7 FEN-1 is implicated in DNA replication and repair in yea

8 FEN-1 is present in the NEIL1 immunocomplex isolated fro

9 FEN-1 mutations altering PCNA binding should reduce acti

10 FEN-1 nucleases have been associated with the base excis

11 FEN-1 proteins are a family of nucleases essential for l

12 FEN-1 recognizes the 5'-flap DNA structure and cleaves i

13 FEN-1 stimulates the activity of NEIL1 in vitro in excis

14 FEN-1 then removes the remaining flap to produce a suita

15 , including the enzymes flap endonuclease 1 (FEN-1) and DNA ligase I that complete the processing and

16 I with the BER factors flap endonuclease 1 (FEN-1) and DNA polymerase delta/epsilon was also observe

17 The interaction between flap endonuclease 1 (FEN-1) and proliferation cell nuclear antigen (PCNA) is

18 onstrated that utilizes flap endonuclease 1 (FEN-1) fused to the Fok1 endonuclease, which shows poten

19 that participates with flap endonuclease 1 (FEN-1) in Okazaki fragment processing.

20 Human flap endonuclease 1 (FEN-1) is a member of the structure-specific endonucleas

21 PCNA binds to flap endonuclease 1 (FEN-1), a structure-specific endonuclease involved in DN

22 cture-specific nuclease Flap Endonuclease 1 (FEN-1), an enzyme that is implicated in the processing o

23 at NEIL1 interacts with flap endonuclease 1 (FEN-1), an essential component of the DNA replication.

24 ase beta (Pol beta) and flap endonuclease 1 (FEN-1).

25 RN which interacts with flap endonuclease 1 (FEN-1).

26 g sites for polymerase, flap endonuclease-1 (FEN-1) and ligase during DNA replication and repair.

27 Flap EndoNuclease-1 (FEN-1) and the processivity factor proliferating cell nu

28 A (RPA), which inhibits flap endonuclease-1 (FEN-1) but stimulates Dna2 nuclease for cleavage.

29 lyses show that WRN and Flap Endonuclease-1 (FEN-1) form a complex in vivo that colocalizes in foci a

30 Flap endonuclease-1 (FEN-1) is a critical enzyme for DNA replication and repa

31 Human flap endonuclease-1 (FEN-1) is a member of the structure-specific endonucleas

32 Human flap endonuclease-1 (FEN-1) is a member of the structure-specific endonucleas

33 Flap endonuclease-1 (FEN-1) is a multifunctional and structure-specific nucle

34 Human flap endonuclease-1 (FEN-1) is a structure-specific endonuclease and exonucle

35 Mammalian flap endonuclease-1 (FEN-1) is a structure-specific metalloenzyme that acts i

36 Flap endonuclease-1 (FEN-1) is a structure-specific nuclease best known for i

37 plication and repair by flap endonuclease-1 (FEN-1) is essential for mammalian genome integrity.

38 gans homologue of human flap endonuclease-1 (FEN-1) that is normally involved in DNA replication and

39 Flap endonuclease-1 (FEN-1), a 43-kDa protein, is a structure-specific and mu

40 e, DNA polymerase beta, flap endonuclease-1 (FEN-1), and PARP-1.

41 nuclease, also known as flap endonuclease-1 (FEN-1).

42 Our results suggest that CRN-1/FEN-1 may play a critical role in switching the state of

43 27p and phage T5 5'-->3' exonuclease (also a FEN-1 homolog).

44 Although a FEN-1 antibody cross-reacting species was detected in th

45 e can, indeed, play a role in expansion by a FEN-1-dependent mechanism.

46 Rad2, a FEN-1 flap endonuclease homolog, is involved in processi

47 to physically bind and stimulate acetylated FEN-1 cleavage activity to the same extent as unacetylat

48 After FEN-1/RTH-1 action and extension by polymerization, DNA

49 Although FEN-1/RTH-1 nuclease often requires an upstream primer f

50 PARP-1 and FEN-1, therefore, cooperate to activate long patch BER.

51 tween proliferating cell nuclear antigen and FEN-1.

52 primary binding sites for both Pol beta and FEN-1 mapped to the TRF2 NH2-terminal and COOH-terminal

53 BLM and FEN-1 are associated with each other in human nuclei as

54 role of the mammalian nucleases RNase HI and FEN-1 in RNA primer removal has been substantiated by se

55 for studying the homologues of RNase HI and FEN-1, i.e., RNase H(35) and Rad27p, respectively.

56 When both ligase and FEN-1/RTH-1 were present simultaneously, some of the 5'-

57 Interaction between NEIL1 and FEN-1 is essential for efficient NEIL1-initiated LP-BER.

58 Furthermore, mutant Rad27 and FEN-1 enzymes with partial flap endonuclease activity bu

59 ion of RNase HI, cutting on the 5'-side, and FEN-1/RTH-1 nuclease, cleaving on the 3'-side.

60 ed light on the aspects of DNA structure and FEN-1 DNA-binding elements that affect substrate cleavag

61 that the two enzymatic activities, UVDE and FEN-1, are part of an alternative excision repair pathwa

62 evidence for an interaction between WRN and FEN-1 in vivo and suggest that these proteins function t

63 A physical interaction between WRN and FEN-1 is demonstrated by their co-immunoprecipitation fr

64 evidence for the interaction between WRN and FEN-1.

65 rther the corresponding interactions between FEN-1 residues and DNA substrates, we performed biochemi

66 The BLM-FEN-1 physical interaction is mediated through a region

67 continuously the kinetics of DNA cleavage by FEN-1 and to separate experimentally the binding and cat

68 s necessary for both binding and cleavage by FEN-1.

69 rand displacement synthesis) were cleaved by FEN-1.

70 sis (long patch repair (LP-BER)) mediated by FEN-1 and stimulated by PCNA.

71 We report here that faulty processing by FEN-1 initiates repeat instability in mammalian cells.

72 activity exist within a single enzyme called FEN-1 [flap endo-nuclease and 5(five)'-exo-nuclease].

73 The conserved FEN-1 C terminus binds proliferating cell nuclear antige

74 a reconstituted BER assay system containing FEN-1, omission of PCNA caused the accumulation of pre-e

75 In contrast, FEN-1 requires a free single-stranded 5' terminus and ap

76 The deduced FEN-1 protein sequences share approximately 75% similari

77 in DNA repair, exogenous nuclease-defective FEN-1 causes repeat instability and aberrant DNA repair.

78 human cells expressing a nuclease-defective FEN-1, we provide direct evidence that an unprocessed FE

79 r more FEN-1 activities presumably to direct FEN-1 to a particular DNA metabolic pathway.

80 27 deletion, which eliminates the endogenous FEN-1 gene product.

81 cised by the structure-specific endonuclease FEN-1 and approximately 2-8 nucleotides are incorporated

82 and the DNA structure-specific endonuclease FEN-1(Rad27) in the processing of DNA ends to be joined

83 y that is provided by the flap endonuclease (FEN-1) in the nucleus, resulting in multinucleotide repa

84 Flap endonuclease (FEN-1) removes 5' overhanging flaps in DNA repair and pr

85 ctures that inhibit human flap endonuclease (FEN-1).

86 bstrates for cleavage by flap endonucleases (FEN-1 proteins).

87 nding and catalytic mechanisms of the entire FEN-1 family of structure-specific nucleases.

88 pus laevis homologue of the endo/exonuclease FEN-1 (DNase IV) have been cloned using a polymerase cha

89 uman 5' flap endonuclease/5'-3' exonuclease (FEN-1), a DNA structure-specific nuclease implicated in

90 uman 5' flap endonuclease/5'-3' exonuclease (FEN-1), a DNA structure-specific nuclease implicated in

91 uman 5'-flap endonuclease/5'-3' exonuclease (FEN-1), a genome stability factor involved in Okazaki fr

92 This 42 kDa endo-/exonuclease, FEN-1, is highly homologous to human XP-G, Saccharomyces

93 WRN was shown to facilitate FEN-1 binding to its preferred double-flap substrate thr

94 The ability of WRN to facilitate FEN-1 cleavage of DNA replication/repair intermediates m

95 n Huntington's disease mice heterozygous for FEN-1 display a tendency toward expansions over contract

96 Structural requirements for FEN-1 and PCNA loading provide an interesting picture of

97 ormation pertinent to nuclease activity from FEN-1, the D181A mutant, the wild-type FEN-1.34-mer DNA

98 The crystal structure of Pyrococcus furiosus FEN-1, active-site metal ions, and mutational informatio

99 How FEN-1 is involved in multiple pathways, of which some ar

100 a substrate-binding model that explains how FEN-1, which has a single active center, can have seemin

101 However, FEN-1 is not efficient in cleaving the short flap, and w

102 We have cloned a human FEN-1 gene, overexpressed it in Escherichia coli, purifi

103 no acid residues, Arg-47 and Arg-70 in human FEN-1, as candidates responsible for substrate binding.

104 through Lys-345) in the C terminus of human FEN-1 (hFEN-1) was shown to be responsible for the inter

105 e identified a 28-amino acid region of human FEN-1 (residues 328-355) and a 29-amino acid region of h

106 show that the C-terminal extension of human FEN-1 likely interacts with the downstream duplex portio

107 ive site residues Glu160 and Asp181 of human FEN-1 nuclease in binding and catalysis of the flap DNA

108 made by Rad27p, the yeast homologue of human FEN-1 protein.

109 approximately 75% similarity with the human FEN-1 nuclease in the conserved nuclease domains, and ex

110 propose a reasonable model for how the human FEN-1 protein interacts with its DNA substrates.

111 Using human FEN-1 in this study, we identified two positively charge

112 olymerases, physically associates with human FEN-1 and stimulates its endonucleolytic activity at bra

113 nd how Mg2+ and flap DNA interact with human FEN-1.

114 Eight restrictively conserved amino acids in FEN-1 have been converted individually to an alanine to

115 e LP-BER activity was marginally affected in FEN-1-depleted mitochondrial extracts, further supportin

116 A conserved arginine in FEN-1 (Arg339) and XPG (Arg992) was found to be crucial

117 dering of unstructured C-terminal regions in FEN-1 and PCNA creates an intermolecular beta sheet inte

118 exonucleases and endonucleases that includes FEN-1, XPG, and GEN1.

119 structure suggests that DNA binding induces FEN-1 to clamp onto the cleavage junction to form the pr

120 d the mammalian homolog of yFEN-1 (DNase IV, FEN-1, or MF1) participates in Okazaki fragment maturati

121 ction is probably important for PCNA to load FEN-1 to the replication fork, to coordinate the sequent

122 Both are 80% identical to mammalian FEN-1 proteins and 55% identical to the yeast homologues

123 m distinct complexes that affect one or more FEN-1 activities presumably to direct FEN-1 to a particu

124 ze the interaction between Mg(2+) and murine FEN-1 (mFEN-1).

125 The mutant FEN-1 exhibited one-tenth the specific activity of wild

126 t Arg(47), Arg(70), and Lys(326)-Arg(327) of FEN-1 interact with the upstream duplex of DNA substrate

127 mediated by the motif (337)QGRLDDFFK(345) of FEN-1, such that an F343AF344A (FFAA) mutant cannot bind

128 An understanding of the ability of FEN-1 to recognize and bind a flap DNA substrate is crit

129 and gap-dependent endonuclease activities of FEN-1 play a role in the resolution of secondary structu

130 ytic and exonucleolytic cleavage activity of FEN-1 and this functional interaction is independent of

131 el explaining the exonucleolytic activity of FEN-1 in terms of its endonucleolytic activity is propos

132 The nuclease activity of FEN-1 is essential for both DNA replication and repair.

133 he reaction was initiated by the addition of FEN-1, the cleavage kinetics were dependent on enzyme co

134 To clarify the molecular basis of FEN-1 specificity and PCNA activation, we report here st

135 n(2+), are required for the active center of FEN-1 nucleases.

136 WRN enhanced the efficiency of FEN-1 cleavage rather than DNA substrate binding.

137 ucture can defeat the protective function of FEN-1, leading to site-specific expansions.

138 k, to coordinate the sequential functions of FEN-1 and other enzymes, and to stimulate its enzyme act

139 led to bind to the scRad27 (yeast homolog of FEN-1) nuclease.

140 ctural changes induced by the interaction of FEN-1 with substrate DNA and Mg2+.

141 nd reveal differences in the interactions of FEN-1 and Cdc9 with the two PCNA interfaces that may con

142 The interacting interface of FEN-1 is localized in its disordered C-terminal region u

143 idues completely blocked the localization of FEN-1 into the nucleus, whereas mutagenesis of the KKK c

144 To further study this effect, a mutant of FEN-1 was identified that retained full nuclease activit

145 By using DNA binding deficient mutants of FEN-1, we determine that the GEN activity is analogous t

146 series of binding-deficient point mutants of FEN-1.

147 stimulation is dependent on the presence of FEN-1.

148 into the chromosome, despite the presence of FEN-1/RTH-1 nuclease.

149 in (WRN) dramatically stimulates the rate of FEN-1 cleavage of a 5' flap DNA substrate.

150 ubstrates interacts with the clamp region of FEN-1.

151 nks adjacent PCNA and DNA binding regions of FEN-1 and suggests how PCNA stimulates FEN-1 activity.

152 that is adjacent to the PCNA binding site of FEN-1.

153 cterize the mechanism for WRN stimulation of FEN-1 cleavage, we have determined the effect of WRN on

154 ng the appropriate relative stoichiometry of FEN-1 and DNA ligase I, which compete for binding to pro

155 eveal a change in the secondary structure of FEN-1 induced by substrate DNA binding.

156 ge is observed in the secondary structure of FEN-1 upon Mg2+ binding to the wild type or to the noncl

157 CNA activation, we report here structures of FEN-1:DNA and PCNA:FEN-1-peptide complexes, along with f

158 res that are the physiological substrates of FEN-1 during replication.

159 the extreme C-terminal 18 amino acid tail of FEN-1 that is adjacent to the PCNA binding site of FEN-1

160 have truncated or mutated the C-terminus of FEN-1 to identify amino acid residues that are critical

161 with DNA substrates were discussed based on FEN-1 cleavage patterns using different substrates.

162 of WRN and PCNA and their combined effect on FEN-1 nuclease activity suggest that they may coordinate

163 ication, we have tested the effect of WRN on FEN-1 cleavage of several DNA substrate intermediates th

164 Flap endonuclease-1 or FEN-1 is a structure-specific and multifunctional nuclea

165 nteractions might be regulated with the PCNA-FEN-1 interaction during DNA replication and repair.

166 The importance of the PCNA-FEN-1 interaction in BER was investigated.

167 report here structures of FEN-1:DNA and PCNA:FEN-1-peptide complexes, along with fluorescence resonan

168 nd character-ization of the putative S.pombe FEN-1 homolog, Rad2p.

169 se activity of the Schizosaccharomyces pombe FEN-1 protein Rad2p requires a 5'-phosphoryl moiety to e

170 ng cell nuclear antigen (PCNA) and positions FEN-1 to act primarily as an exonuclease in DNA replicat

171 cture may actively participate by preventing FEN-1 cleavage of displaced Okazaki fragments.

172 had been previously classified as a putative FEN-1 protein based on amino acid homology, there has be

173 ved to any detectable level, deleting RAD27 (FEN-1 of yeast) leads to a 4.4-fold reduction specifical

174 sts that the endonuclease activity of Rad27 (FEN-1) plays a role in limiting recombination between sh

175 these genes belong to the Rad2/XPG and Rad27/FEN-1 families, which are structure-specific nucleases f

176 Because Rad27/FEN-1 acts specifically at 5' flap structures, these res

177 Recently, FEN-1 has been reported to also possess a gap endonuclea

178 th catalytically active purified recombinant FEN-1 deletion mutant proteins that lack either the WRN/

179 dentify cellular factors that might regulate FEN-1 activity.

180 t, a different E3 ubiquitin ligase regulates FEN-1 turnover.

181 tide excision repair, which does not require FEN-1 or PCNA.

182 not require a free upstream end to stimulate FEN-1 cleavage of the 5' flap substrate.

183 ignificantly the ability of WRN to stimulate FEN-1 incision activities.

184 WRN effectively stimulated FEN-1 cleavage on a flap DNA substrate with streptavidin

185 WRN stimulated FEN-1 cleavage of flap substrates with a terminal monori

186 a regressed replication fork and stimulates FEN-1 to cleave the unwound product in a structure-depen

187 e C-terminal domain of WRN or BLM stimulates FEN-1 cleavage of its proposed physiological substrates

188 WRN effectively stimulates FEN-1 cleavage of branch-migrating double-flap structure

189 ns of FEN-1 and suggests how PCNA stimulates FEN-1 activity.

190 te that the mechanism whereby WRN stimulates FEN-1 cleavage is distinct from that proposed for the fu

191 nal changes and coordination with subsequent FEN-1 activity.

192 quire nucleosome remodeling in vivo and that FEN 1 activity during Okazaki fragment processing can oc

193 In this study, we demonstrated that FEN-1(Rad27) physically and functionally interacted with

194 Our data indicate that FEN-1 and the D181A mutant each have a radius of gyratio

195 There is much evidence to indicate that FEN-1 efficiently cleaves single-stranded DNA flaps but

196 ays form stable secondary structures or that FEN-1 has an alternative approach to resolve the seconda

197 The FEN-1/PCNA interaction is mediated by the motif (337)QGR

198 d the borders of the interaction between the FEN-1 protein and its DNA substrates and identified six

199 mutations, such as rad27 Delta, encoding the FEN-1 nuclease involved in several aspects of genomic st

200 complex present, we estimated the Kd for the FEN-1-flap DNA substrate to be 7.5 nM in the absence of

201 d the crystallographic ainformation from the FEN-1.DNA complex that we published recently we are able

202 In the absence of magnesium, the FEN-1.34-mer DNA flap complex has an Rg value of approxi

203 f WRN and BLM with FEN-1, we have mapped the FEN-1 binding site on the two RecQ helicases.

204 fect of WRN on the kinetic parameters of the FEN-1 cleavage reaction.

205 ite through site-directed mutagenesis of the FEN-1 gene.

206 XPG is a member of the FEN-1 structure-specific endonuclease family.

207 d2 family and they cluster with genes of the FEN-1 subfamily, which are known to be involved in DNA r

208 This result indicates the orientation of the FEN-1-DNA interaction.

209 les arising from magnesium activation of the FEN-1.34-mer DNA flap complex is consistent with the pro

210 , our data demonstrate the importance of the FEN-1/PCNA complex in DNA replication and in the embryon

211 To determine the physiological roles of the FEN-1/PCNA interaction in a mammalian system, we knocked

212 ate through its protein interaction with the FEN-1 C-terminal binding site.

213 of the WRN/BLM physical interaction with the FEN-1 C-terminal tail was confirmed by functional intera

214 cna-90 was defective in interaction with the FEN-1 endo-exonuclease (RTH1 product).

215 erminal domain that shares homology with the FEN-1 interaction domain of the Werner syndrome protein,

216 Thus, FEN-1 enzymes and likely reaction mechanisms are conserv

217 ntrast, a PCNA mutant that could not bind to FEN-1 was unable to stimulate excision.

218 and sufficient for high affinity binding to FEN-1 (K(D) approximately = 0.2 microm).

219 PCNA binds to FEN-1 and stimulates its nuclease activity, but the phys

220 ic repair endonuclease that is homologous to FEN-1.

221 handoff of intermediates from polymerase to FEN-1 to ligase during DNA replication and repair.

222 , particularly one that is made resistant to FEN-1/RTH-1-directed cleavage by extension of an inhibit

223 A gene with sequence similarity to FEN-1 protein-encoding genes, rad2 +, has been identifie

224 one-tenth the specific activity of wild type FEN-1 in the reconstituted BER assay, and this repair de

225 from FEN-1, the D181A mutant, the wild-type FEN-1.34-mer DNA flap complex, and the D181A.34-mer DNA

226 activity to the same extent as unacetylated FEN-1.

227 provide direct evidence that an unprocessed FEN-1 substrate is a precursor to instability.

228 However, when FEN-1 is absent from the cell, alternative pathways to s

229 suggest that they may coordinately act with FEN-1.

230 nding of the interaction of WRN and BLM with FEN-1, we have mapped the FEN-1 binding site on the two

231 hnRNP A1), which forms a direct complex with FEN-1 and stimulates its enzymatic activities.

232 he interaction of WRN and BLM helicases with FEN-1, and how these interactions might be regulated wit

233 cna-79 (IL126,128AA) failed to interact with FEN-1, but, surprisingly, pcna-79 was still very active

234 least 20 proteins are known to interact with FEN-1; some form distinct complexes that affect one or m

235 long-patch BER through its interaction with FEN-1.

236 The WRN-FEN-1 functional interaction is independent of WRN catal

237 llectively, the results suggest that the WRN-FEN-1 interaction is biologically important in DNA metab

238 derstand the potential importance of the WRN-FEN-1(1) interaction in DNA replication, we have tested

239 Employing a yeast FEN-1 mutant, E176A, which is deficient in exonuclease (

240 WRN and yeast FEN-1 were reciprocally co-immunoprecipitated from extra

241 A physical interaction between yeast FEN-1 and WRN is demonstrated by yeast FEN-1 affinity pu

242 yeast FEN-1 and WRN is demonstrated by yeast FEN-1 affinity pull-down experiments using transformed d

243 We now show that the yeast FEN-1 (yFEN-1) nuclease interacts genetically and bioche