ライフサイエンスコーパス: Pol beta

1 Pol beta activity is enhanced by flap endonuclease (FEN1

2 Pol beta also misinserts a purine nucleotide opposite th

3 Pol beta also was able to perform intersegmental transfe

4 Pol beta is also widely used as a model polymerase for s

5 Pol beta is mutated in a large number of colorectal tumo

6 Pol beta localizes to synapsed axes during zygonema and

7 Pol beta promoted incorporation of dCMP and dAMP, along

8 Pol beta-deficient spermatocytes yielded reduced steady-

9 Pol-beta-deficient cells were significantly more suscept

10 a molecule with the active site of the 1 : 1 Pol beta-DNA complex, while the latter demonstrates how

11 -dimensional structural models for the 2 : 1 Pol beta-DNA and 1 : 1 Pol X-DNA complexes were generate

12 plexes, we propose that the asymmetric 2 : 1 Pol beta-DNA complex enhances the function of Pol beta.

13 The results indicated formation of a 2 : 1 Pol beta-DNA complex, whereas only 1 : 1 Pol X-DNA compl

14 at, when repair synthesis proceeds through a Pol beta-dependent single nucleotide replacement mechani

15 lly those with dRP lyase activity, we used a Pol beta null cell extract and BER intermediate as bait

16 2 that results from formation of an abortive Pol beta-gapped DNA-dATP complex is consistent with an o

17 A second group of polymerases (Pol alpha, Pol beta, and T7(-)) fails to extend all non-H-bonding b

18 utations are further correlated with altered Pol beta enzymatic activity.

19 DNA binding specificity inherent in APE and Pol beta, although coordination also may be facilitated

20 and Pol iota (family Y), and Pol lambda and Pol beta (family X).

21 te in cell extracts prepared from normal and Pol beta-null mouse cells and show that the reduced repa

22 S. cerevisiae whereas expression of another Pol beta dominant negative mutant, Pol beta-TR, does not

23 coordination also may be facilitated by APE.Pol beta.DNA ternary complex formation with appropriate

24 zyme concentrations a ternary complex of APE.Pol beta.DNA that formed specifically at BER intermediat

25 alysis suggests that these motions allow apo Pol beta to sample a conformation similar to the gapped

26 In apo Pol beta, molecular motions, primarily isolated to the D

27 ciency in mitochondrial extracts from APTX-/-Pol beta-/- cells.

28 at they follow the same kinetic mechanism as Pol beta, while differing in relative rates of single-nu

29 me-DNA complexes of rat DNA polymerase beta (Pol beta) and African swine fever virus DNA polymerase X

30 of DNA polymerases: rat DNA polymerase beta (Pol beta) and African swine fever virus DNA polymerase X

31 t TRF2 physically bound DNA polymerase beta (Pol beta) and flap endonuclease 1 (FEN-1).

32 and also interacts with DNA polymerase beta (Pol beta) and other base excision repair (BER) proteins.

33 cised abasic site, both DNA polymerase beta (Pol beta) and the DNA ligase IIIalpha-XRCC1 heterodimer

34 DNA polymerase beta (Pol beta) carries out strand displacement synthesis, fol

35 in RNA-mediated loss of DNA polymerase beta (Pol beta) expression in human breast cancer cells increa

36 topped-flow analyses of DNA polymerase beta (Pol beta) had provided important mechanistic insight, th

37 eotide incorporation by DNA polymerase beta (Pol beta) has been well-studied, a true understanding of

38 excision repair enzyme DNA polymerase beta (Pol beta) in complex with a 1-nt gapped DNA substrate co

39 ate states of mammalian DNA polymerase beta (Pol beta) in its wild-type and an error-prone variant, I

40 negative mutants of rat DNA polymerase beta (Pol beta) interfere with repair of alkylation damage in

41 DNA polymerase beta (Pol beta) is a 39-kDa enzyme that performs the vital cel

42 During DNA repair, DNA polymerase beta (Pol beta) is a highly dynamic enzyme that is able to sel

43 DNA polymerase beta (Pol beta) is a key enzyme in DNA base excision repair, a

44 sphate (dRP) removal by DNA polymerase beta (Pol beta) is a pivotal step in base excision repair (BER

45 rminal domain (NTD) and DNA polymerase beta (Pol beta) is central to base excision repair of damaged

46 DNA polymerase beta (Pol beta) is one of the key enzymes in the base excision

47 DNA polymerase beta (Pol beta) is responsible for the repair synthesis in thi

48 formational step in the DNA polymerase beta (Pol beta) kinetic pathway.

49 have shown earlier that DNA polymerase beta (Pol beta) localizes to the synaptonemal complex (SC) dur

50 DNA polymerase beta (Pol beta) plays a key role in base excision repair (BER)

51 In contrast, DNA polymerase beta (Pol beta) synthesis is inhibited in a major fraction ( a

52 DNA polymerase beta (Pol beta) was implicated recently as the major polymeras

53 DNA polymerase beta (Pol beta), a key enzyme in the DNA base excision repair

54 DNA polymerase beta (Pol beta), a member of the DNA polymerase X family that

55 ibly inhibits repair by DNA polymerase beta (Pol beta), an integral enzyme in base-excision repair.

56 erminal 8 kDa domain of DNA polymerase beta (Pol beta), and for the homologous domain of DNA polymera

57 The DNA repair enzyme, DNA polymerase beta (Pol beta), is among the most discriminating, being inact

58 e that can ubiquitylate DNA polymerase beta (Pol beta), the major BER DNA polymerase.

59 man cancers overexpress DNA polymerase beta (Pol beta), the major DNA polymerase involved in base exc

60 the catalytic cycle of DNA polymerase beta (Pol beta), using a synthetic DNA primer/template contain

61 ce assays for mammalian DNA polymerase beta (Pol beta), we have previously identified a fast fluoresc

62 endonuclease (APE), and DNA polymerase beta (Pol beta), which catalyze the first three steps in BER,

63 ndonuclease 1 (APE) and DNA polymerase beta (Pol beta).

64 he AP lyase activity of DNA polymerase beta (Pol beta).

65 avily reliant upon host DNA polymerase beta (Pol beta).

66 that APC interacts with DNA polymerase beta (Pol-beta) and flap endonuclease 1 (Fen-1) and blocks Pol

67 itution also affects the interaction between Pol beta and proliferating cell nuclear antigen (PCNA).

68 more similar to those observed in the binary Pol beta-dATP complex.

69 ) and flap endonuclease 1 (Fen-1) and blocks Pol-beta-directed strand-displacement synthesis.

70 The primary binding sites for both Pol beta and FEN-1 mapped to the TRF2 NH2-terminal and C

71 our findings indicate that APC blocked both Pol-beta-directed SN- and LP-BER pathways and increased

72 we show that removal of the 5'-dRP block by Pol beta is unaffected by NCP constraints at all sites t

73 s to play a minimal role in the catalysis by Pol beta.

74 t these regions can be scanned for damage by Pol beta.

75 mechanisms of damage search and location by Pol beta are largely unknown, but are critical for under

76 s leads to formation of the product complex, Pol beta-DNA-Cr(III).PCP, whose structure is also report

77 Here, we constructed Pol beta R137Q knock-in mice, and found that homozygous

78 r stability of newly synthesized cytoplasmic Pol beta that is used as a source for nuclear Pol beta i

79 excision repair (BER) proteins XRCC1 and DNA Pol beta, adding a new level of regulation for BER.

80 t following incision by AP endonuclease, DNA Pol beta recognizes and binds to the incised abasic site

81 man tumors characterized to date express DNA Pol beta variants, many of which result from a single nu

82 ence for a stable preexisting complex of DNA Pol beta with the DNA ligase IIIalpha-XRCC1 heterodimer.

83 W1 and ISW2 from yeast significantly enhance Pol beta accessibility to the refractory nicked sites in

84 approach we find that in human cell extracts Pol beta is the major DNA polymerase incorporating the f

85 orporation into damaged DNA is essential for Pol beta repair function, and several studies have impli

86 localization patterns reflect a function for Pol beta in recombination and/or synapsis, we used condi

87 on-covalent step not previously reported for Pol beta.

88 lexes, this work further validates that, for Pol beta, fidelity is dictated by the differences in fre

89 of either wild-type human Pol beta or human Pol beta with an inactivating mutation specific to the p

90 by complementation of either wild-type human Pol beta or human Pol beta with an inactivating mutation

91 ed a processive search assay to determine if Pol beta has evolved a mechanism for efficient DNA damag

92 A (RPA), RAD51 and DMC1 are less abundant in Pol beta-deficient spermatocyte nuclei.

93 of a rate-limiting conformational change in Pol beta's reaction pathway.

94 es have implicated conformational changes in Pol beta as a determinant of this repair fidelity.

95 e to the intimate role of protein motions in Pol beta fidelity.

96 rk, the rate constants for domain motions in Pol beta have been determined by solution NMR relaxation

97 Mutations in Pol beta have been linked to various cancers, and these

98 RPase activity, does not stimulate repair in Pol beta-null cells.

99 se cells and show that the reduced repair in Pol beta-null extracts can be complemented by addition o

100 , we have mapped the APC interaction site in Pol-beta and have found that Thr79, Lys81, and Arg83 of

101 l-beta blocked SN-BER activity by inhibiting Pol-beta-directed deoxyribose phosphate lyase activity.

102 iplatin sensitivity and DNA repair involving Pol beta.

103 oxaliplatin treatment compared with isogenic Pol beta-expressing cells.

104 MR studies on [methyl-13C]methionine-labeled Pol beta complexed with double-hairpin DNA, used to mode

105 Mutant Pol beta deficient in DNA synthesis, but with normal dRP

106 ition of purified wild type but not a mutant Pol beta protein that does not interact with the DNA lig

107 to WT Pol beta, suggesting that this mutant Pol beta can be used to incorporate Rp-dNTPalphaS into D

108 f another Pol beta dominant negative mutant, Pol beta-TR, does not.

109 e of the Pol beta dominant negative mutants, Pol beta-14, increases the spontaneous mutation rate of

110 mentation assay, we demonstrate that mutated Pol beta without dRPase activity is able to stimulate lo

111 static relationship of the dominant negative Pol beta mutants to genes known to be involved in repair

112 milar in fold to those of other noncanonical Pol beta RNA polymerases, and the two zinc knuckle motif

113 ol beta that is used as a source for nuclear Pol beta involved in DNA repair.

114 with the intersegmental searching ability of Pol beta being at least 6- and approximately 2-fold high

115 ths, the intramolecular searching ability of Pol beta is at least 4-fold higher than that of Pol mu a

116 A knockdown of Mule leads to accumulation of Pol beta and increased DNA repair.

117 t the dRPase and DNA synthesis activities of Pol beta are coupled and that both of these Pol beta fun

118 he 5'-deoxyribosephosphate lyase activity of Pol beta is essential for repair of both lesions.

119 esions, and that the 5'dRP lyase activity of Pol beta is required to restore resistance to temozolomi

120 synapsis defects and Prophase I apoptosis of Pol beta-deficient spermatocytes are likely a direct con

121 se defects impair the DNA repair capacity of Pol beta in reconstitution assays, as well as in cellula

122 The amino-terminal 8 kDa domain of Pol beta has an activity for excising a 5'-deoxyribose p

123 ing a novel function for the lyase domain of Pol beta.

124 o to the significantly reduced expression of Pol beta.

125 our standing hypothesis that the fidelity of Pol beta originates from destabilization of the mismatch

126 se data suggest that the repair footprint of Pol beta mainly resides within accessible regions of the

127 the apo and substrate-bound, binary forms of Pol beta.

128 ol beta-DNA complex enhances the function of Pol beta.

129 Irreversible inhibition of Pol beta by DOB suggests that this lesion provides a che

130 the design of a new family of inhibitors of Pol beta.

131 OR cells, small interfering RNA knockdown of Pol beta delayed the repair of oxaliplatin-induced DNA d

132 te its biological significance, knowledge of Pol beta activities on NCPs is still limited.

133 st an alteration in the dynamic landscape of Pol beta due to substrate binding.

134 We further show that the cellular levels of Pol beta and its ubiquitylated derivative are modulated

135 found to have higher constitutive levels of Pol beta protein, faster in vitro repair of a DNA substr

136 ubiquitylated Pol beta, decreased levels of Pol beta, and a subsequent deficiency in BER, leading to

137 Localization of Pol beta to meiotic chromosomes requires the formation o

138 r data suggest that the kinetic mechanism of Pol beta is not an exception among polymerases, and furt

139 ovide useful information to the mechanism of Pol beta.

140 w fluorescence change occurs after mixing of Pol beta.DNA.Rh(III)dNTP with Mg(II).

141 neither dRPase nor DNA synthesis mutants of Pol beta alone, or the two together, were able to comple

142 e-limiting in the overall kinetic pathway of Pol beta, yet is kinetically significant in subdomain re

143 for mutagenesis to occur in the presence of Pol beta-14 but that it is not merely the presence of a

144 ymerases to answer whether the properties of Pol beta are exceptional or general.

145 d to be associated with the stabilization of Pol beta protein on chromatin.

146 ed on the basis of the tertiary structure of Pol beta and the corresponding regions of Pol X were inf

147 tent with recent crystallographic studies of Pol beta complexed with 5-phosphorylated abasic sugar an

148 three probes in the stopped-flow studies of Pol beta to obtain new, direct evidence for our previous

149 , enhanced both its own activity and that of Pol beta on nucleosome substrates.

150 on in this cell type because the Km value of Pol beta is much higher than the dNTP concentrations fou

151 d have found that Thr79, Lys81, and Arg83 of Pol-beta were critical for its interaction with APC.

152 One Pol beta mutation that has been identified in tumors, R1

153 tuated within the middle of the BER pathway, Pol beta must efficiently locate its substrates before d

154 biquitylation of the key BER DNA polymerase (Pol beta) and demonstrate that USP47 is required for sta

155 f the stereoselectivity of a DNA polymerase, Pol beta from rat brain, toward Rp and Sp isomers of dAT

156 Three human DNA polymerases (Pol beta, Pol delta and Pol epsilon) have been proposed

157 amino acid site that is essential for proper Pol beta functioning in maintaining genomic stability an

158 can be complemented by addition of purified Pol beta.

159 We demonstrate that the rat Pol beta mutants interfere with the base excision repair

160 the 8 kDa 5'-dRP lyase domain of the second Pol beta molecule with the active site of the 1 : 1 Pol

161 well as from the determination that soaking Pol beta crystals with Mn(II) ions leads to formation of

162 ished to determine whether the S229L somatic Pol beta variant identified in a stage 3 colorectal tumo

163 TRF2 also stimulated Pol beta strand displacement DNA synthesis in reconstitu

164 TRF2 stimulated Pol beta primer extension DNA synthesis on telomeric and

165 omplex was associated with slightly stronger Pol beta gap-filling and much stronger 5'-deoxyribose ph

166 Consistent with previous studies, Pol beta-deficient mouse fibroblasts were not hypersensi

167 In a different model system, Pol beta-deficient fibroblasts were less able to repair

168 ncentrations in whole cell extracts and that Pol beta has a 7-fold higher affinity to the incised aba

169 We further demonstrate that Pol beta and the DNA ligase IIIalpha-XRCC1 heterodimer a

170 They demonstrate that Pol beta modulates the sensitivity of cells to oxaliplat

171 We find that Pol beta-deficient spermatocytes are defective in meioti

172 These results imply that Pol beta-dependent 5'dRP lyase activity is the rate-limi

173 ether, these findings strongly indicate that Pol beta is required at a very early step in the process

174 These results indicate that Pol beta-dependent base excision repair of uracil opposi

175 rsists on meiotic chromatin, indicating that Pol beta is critical for the repair of SPO11-induced dou

176 into nucleosome core particles revealed that Pol beta is not processive in the context of a nucleosom

177 These assays revealed that Pol beta scans DNA using a processive hopping mechanism

178 ed guanine was dramatically decreased at the Pol beta insertion step, but not blocked.

179 tion repair pathway that is disrupted by the Pol beta dominant negative mutants, we studied the epist

180 eterodimer was decreased about 2-fold in the Pol beta-deficient cell extract but was rescued by addit

181 In addition, expression of one of the Pol beta dominant negative mutants, Pol beta-14, increas

182 RET)-based system to monitor movement of the Pol beta fingers domain during catalysis in the presence

183 Expression of the Pol beta-14 mutant in cells lacking APN1 activity does n

184 PCNA could promote the Pol beta-dependent long-patch pathway by stimulation of

185 of predicted structural similarity with the Pol beta superfamily as critical for Trf4p/Pol sigma's e

186 lyase activities than was observed with the Pol beta.DNA binary complex.

187 The Pol-beta protein (T79A/K81A/R83A) blocked strand-displac

188 Pol beta are coupled and that both of these Pol beta functions are essential during short patch BER

189 cks in linker DNA are far more accessible to Pol beta in highly folded oligonucleosomes.

190 Expression of wild-type Pol beta in pol beta(-/-) mouse embryonic fibroblast (ME

191 7 causes an increased level of ubiquitylated Pol beta, decreased levels of Pol beta, and a subsequent

192 elation to the crystal structures of various Pol beta complexes, as well as previous steady-state kin

193 cesses, including base excision repair where Pol beta catalyzes two key enzymatic steps: 5'-dRP lyase

194 We examined whether Pol beta, expressed at levels found in tumor cells, is i

195 wo crystal forms of XRCC1-NTD complexed with Pol beta have been solved, revealing that the XRCC1-NTD

196 IIIalpha complex that does not interact with Pol beta and tested their activities in BER reconstitute

197 g XRCC1 protein, that does not interact with Pol beta has reduced ability to rescue the hydrogen pero

198 n which disrupts functional interaction with Pol beta, affected the ligation efficiency of the mutant

199 XRCC1-NTD forms additional interactions with Pol beta, enhancing affinity by an order of magnitude.

200 merase active site, and superimposition with Pol beta in a ternary complex suggests a shift in the po

201 R indicated that the interaction of APC with Pol-beta blocked SN-BER activity by inhibiting Pol-beta-

202 activity toward Rp-dATPalphaS relative to WT Pol beta, suggesting that this mutant Pol beta can be us

203 anism of misincorporation for wild-type (WT) Pol beta and an error-prone I260Q variant using stopped-

204 data suggest an important role for the XRCC1-Pol beta interaction for coordinating the efficiency of