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 olymerase delta (POLd), flap endonuclease 1 (FEN-1), and DNA ligase 1 (LIG1).

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

44 27p and phage T5 5'-->3' exonuclease (also a FEN-1 homolog).

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

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

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

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

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

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

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

52 tween proliferating cell nuclear antigen and FEN-1.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

81 27 deletion, which eliminates the endogenous FEN-1 gene product.

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

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

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

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

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

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

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

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

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 DNA structure-specific nuclease implicated in

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

165 Only FEN-1 and LIG1 are required for the repair of the minus

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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