ライフサイエンスコーパス: primase

1 e interface in the NTD of DnaB that contacts primase.

2 nto the mechanism of nucleotide synthesis by primase.

3 (p180DeltaN-p70) inhibited RNA synthesis by primase.

4 t a new primer synthesized downstream by the primase.

5 lves interactions of these proteins with DNA primase.

6 hat of a functional primer synthesized by T7 primase.

7 interaction of the yeast ortholog pol1 with primase.

8 ymerase requires RNA primers produced by DNA primase.

9 tacting and sequestering the relaxase-linked primase.

10 on due to impaired binding to both ssDNA and primase.

11 s to elevated RNA primer synthesis by T7 DNA primase.

12 bility of this DNA helicase to interact with primase.

13 ructure of the iron-sulfur cluster domain of primase.

14 occluding this region from interacting with primase.

15 inding activity of D5, the poxvirus helicase-primase.

16 Hence, DnaC controls the access of DnaB by primase.

17 main not found in the archaeal and bacterial primases.

18 g mutated, truncated, chimeric and wild-type primases.

19 priming of DNA templates by enzymes known as primases.

20 and eukaryotes requires the activity of DNA primase, a DNA-dependent RNA polymerase that lays short

21 all organisms depends on the activity of DNA primase, a DNA-dependent RNA polymerase that synthesizes

22 CST/AAF, a DNA polalpha.primase accessory factor, binds POT1b and shortens the e

23 The molecules were docked to the primase active site using the available primase crystal

24 iously unrecognized as critical to the human primase active site.

25 been reported to stimulate the helicase and primase activities of the complex in the presence of ICP

26 The helicase and primase activities of the hexameric ring-shaped T7 gp4 p

27 expresses full and functional helicase (and primase) activities when bound to a gp61 primase subunit

28 ase activity, it substantially augments both primase activity and primase-to-polymerase switching.

29 Furthermore, we show that primase activity exhibits a cooperative dependence on pr

30 Biochemical assays measuring primase activity have been limited to monitoring formati

31 additionally report that although PrimPol's primase activity is required to restore wild-type replic

32 We find that primase activity lowers replisome processivity but only

33 r Mn(II) over Mg(II), suggesting that T7 DNA primase activity modulation when bound to Mn(II) is base

34 sis reveals that RPA serves to stimulate the primase activity of PrimPol.

35 Taken together, these data suggest that the primase activity of the helicase-primase requires format

36 the discontinuous nature of DNA replication, primase activity on the lagging strand is required throu

37 e N-terminal domain in T7 gp4 contains a DNA primase activity, this function is lost in metazoan mtDN

38 To identify mechanisms regulating primase activity, we characterized primase initiation sp

39 th the second subunit (p180C-p70) stimulated primase activity, whereas the whole catalytically active

40 s zinc ions and is essential for maintaining primase activity.

41 he primosome, but significantly inhibits its primase activity.

42 n has helicase activity but no DNA-dependent primase activity.

43 oading on RPA-coated ssDNA and regulation of primase activity.

44 y DNA polymerase-alpha and its intrinsic RNA primase activity.

45 activity, and PrimPol, a DNA polymerase with primase activity.

46 e (PrimPol; CCDC111), an archaeal-eukaryotic primase (AEP) in eukaryotic cells, is involved in chromo

47 is article focuses on the archaeo-eukaryotic primase (AEP) superfamily, drawing on recently character

48 In eukaryotes, a single archaeo-eukaryotic primase (AEP), DNA primase, is required for the initiati

49 Slow primer release from DNA primase allows the polymerase to engage the complex and

50 olecule inhibitors of the activity of T7 DNA primase, an ideal model for bacterial primases due to th

51 s predicted to possess an archaeo-eukaryotic primase and a UL52-like zinc finger domain, the role of

52 lication system of bacteriophage T7 both DNA primase and DNA helicase activities are contained within

53 human primosome, a 340-kilodalton complex of primase and DNA polymerase alpha (Polalpha), synthesizes

54 gging strand synthesis additionally requires primase and DnaE.

55 bserve that signal release is independent of primase and does not seem to require a protein trigger a

56 Disrupting the interaction between primase and helicase in Escherichia coli increases Okaza

57 alled "clamp zones." Loading depends on DnaG primase and is probably driven by Okazaki fragment initi

58 ack of crystal structures of the full-length primase and its complexes with substrates in initiation

59 and is consistent with the requirements for primase and ligase activities as well as earlier electro

60 e ability to disrupt the association between primase and pol alpha allowed us to assess the physiolog

61 The catalytic subunits of primase and pol alpha synthesize composite RNA-DNA prime

62 domains and defining the requirement for its primase and polymerase activities during nuclear DNA rep

63 s, we find that SSBs significantly limit the primase and polymerase activities of PrimPol.

64 that PrimPolY89D has a striking decrease in primase and polymerase activities.

65 een p180C, p70, and p58 regulates the proper primase and polymerase function.

66 PrimPol was recently identified as a TLS primase and polymerase involved in DNA damage tolerance.

67 However, the negative effects of primase and rNTPs on processivity are overcome by the ex

68 New herpesvirus drugs include viral helicase-primase and terminase inhibitors.

69 lication proteins, including DnaA, helicase, primase and the clamp loader, TrfA interaction with the

70 xtension effected by cooperation between DNA primase and the lagging-strand polymerase.

71 nts and binding mode for interactions of DNA primase and thymidylate synthetase (TS) with high and lo

72 l is a recently discovered DNA-dependent DNA primase and translesion synthesis DNA polymerase found i

73 aryotic AEP-like proteins with DNA-dependent primase and/or polymerase activity.

74 is novel assay should be applicable to other primases and inefficient DNA/RNA polymerases, facilitati

75 rations of DNA polymerase-alpha primase (Pol-primase), and the p58 subunit of Pol-primase associates

76 ingle-stranded DNA (ssDNA)-dependent ATPase, primase, and DNA binding activities.

77 and V that also function with the helicase, primase, and sliding clamp in the replisome.

78 ed residues within the ATPase, Topoisomerase/Primase, and Winged helix domains, including four that e

79 n reaction catalyzed by the T. kodakaraensis primase are discussed.

80 olymerase and the zinc-finger domains of DNA primase are involved in the stabilization of the priming

81 that the dramatic conformational changes in primase are necessary to accomplish the initiation and t

82 Primases are crucial RNA polymerases that perform the in

83 Eukaryotic and archaeal primases are heterodimers consisting of small catalytic

84 Eukaryotic and archaeal primases are heterodimers consisting of small catalytic

85 tants A49 and A53 did not interact with UL52 primase as determined by co-immunoprecipitation experime

86 8 of Pol1 abrogates the interaction with the primase, as does mutation to alanine of the invariant am

87 LTA synthases while YvgJ functions as an LTA primase, as indicated by the accumulation of a GroP-Glc(

88 se (Pol-primase), and the p58 subunit of Pol-primase associates with NFIC/CTF1, suggesting that NFI a

89 In eukaryotic cells, the presence of primase at the replication fork is secured by its physic

90 leading Pol epsilon below CMG and Pol alpha-primase at the top of CMG at the replication fork.

91 exponential amplification of genomic DNA in primase-based whole-genome amplification (T4 pWGA).

92 to assess the physiological significance of primase being tethered to the eukaryotic replisome in th

93 lusters using DNA charge transport regulates primase binding to DNA and illustrates chemistry that ma

94 at occur only on the lagging strand, such as primase binding to DnaB helicase, RNA synthesis, and SS

95 revealed that the N-terminal domain of UL52 primase binds UL5 helicase and the middle domain interac

96 r binds weakly to fork DNA in the absence of primase, but forms a much more stable primosome complex

97 ontrols the ability of DnaB to interact with primase by modifying the conformation of the NTD of DnaB

98 Helicase binding to the primase C-terminal helicase-binding domain modulated RNA

99 strate that the [4Fe4S] cluster in human DNA primase can make use of this chemistry to coordinate the

100 DNA primases catalyze the synthesis of oligoribonucleotides

101 DNA primases catalyze the synthesis of the oligoribonucleoti

102 A primase chimera, where PriX is fused to a truncated vers

103 By contrast, the helicase-primase complex (DnaB and DnaG) remains critically assoc

104 and biochemical characterization of the DNA primase complex and its subunits from the archaeon Therm

105 The Polalpha/primase complex assembles the short RNA-DNA fragments fo

106 The DNA polymerase alpha-primase complex forms an essential part of the eukaryoti

107 The DNA Polymerase alpha (Pol alpha)/primase complex initiates DNA synthesis in eukaryotic re

108 The T. kodakaraensis DNA primase complex is a heterodimer containing stoichiometr

109 es simplex virus 1 (HSV-1) UL5/8/52 helicase-primase complex is required for DNA unwinding at the rep

110 The heterotrimeric helicase-primase complex of herpes simplex virus type I (HSV-1),

111 We report that the T. kodakaraensis primase complex preferentially interacts with dNTP rathe

112 he completed RNA-DNA primer by the Pol alpha/primase complex simplifies current models of primer tran

113 pplementation of such reactions with the DNA primase complex supported lagging strand formation as we

114 itelivir, an inhibitor of the viral helicase-primase complex, exhibits antiviral activity in vitro an

115 e latter findings indicate that the archaeal primase complex, in contrast to the eukaryote homolog, c

116 lished primarily by the DNA polymerase alpha-primase complex, which makes the RNA-DNA primers accessi

117 with a completed Okazaki fragment or primer-primase complexes as the recycling mechanism.

118 eriophage DNA replication system that primer-primase complexes have a residence time similar to the t

119 We examined the role of RNA primer-primase complexes left on the lagging ssDNA from primer

120 The collision with primer-primase complexes triggering the early termination of Ok

121 ation machinery includes a trimeric helicase-primase composed of helicase (UL5) and primase (UL52) su

122 Archaea encode a eukaryotic-type primase comprising a catalytic subunit (PriS) and a nonc

123 gging stand synthesis included high helicase-primase concentrations and a lagging strand template who

124 Prokaryotic primases consist of a zinc-binding domain (ZBD) necessar

125 ically characterized the bacterial-like DnaG primase contained within the hyperthermophilic crenarcha

126 In the complex, Pol alpha and primase cooperate in the production of RNA-DNA oligonucl

127 the primase active site using the available primase crystal structure and ranked based on their pred

128 ion of the essential poxvirus virus helicase-primase D5 and show that the active helicase domain of D

129 vealed that some of the molecules inhibit T7 primase-dependent DNA replication.

130 , which promotes polymerase alpha (polalpha)/primase-dependent fill-in throughout the genome and at t

131 activities such as duplex translocation and primase-dependent RNA synthesis.

132 The phage encodes its own primase, DNA ligase, DNA polymerase, and enzymes necessa

133 short RNA-DNA hybrid primers synthesized by primase-DNA polymerase alpha (Prim-Pol alpha) are needed

134 and purified the previously uncharacterized primase DnaG from Mycobacterium tuberculosis (Mtb DnaG).

135 racts with the replicative helicase DnaC and primase DnaG in a ternary complex.

136 Bacterial DNA primase DnaG synthesizes RNA primers required for chromo

137 amage in a reaction that is dependent on the primase, DnaG, but independent of any of the known repli

138 A novel peptide from the DNA primase, DNAP(211-223), was also found.

139 e mirrored by experiments in yeast cells, as primase does not interact in cell extracts with pol1 tha

140 The primase domain and the helicase domain were structurally

141 The N-terminal primase domain of the gene 4 protein of phage T7 compris

142 en the winged helix domain and topoisomerase-primase domain, remote from the DNA.

143 ains a C-terminal helicase and an N-terminal primase domain.

144 n observed between the TOPRIM (topoisomerase/primase) domain and the scissile phosphate.

145 ll-length gp4 contains both the helicase and primase domains.

146 T7 DNA primase, an ideal model for bacterial primases due to their common structural and functional f

147 Primase equilibrates with the fork prior to synthesis of

148 DNA primase facilitates binding of DNA helicase to ssDNA and

149 It tethers Pol alpha and primase, facilitating RNA primer handover from primase t

150 th the bacterial DnaG and archaeo-eukaryotic primase families.

151 primase function and are applicable for DNA primases from other species.

152 rovide notable insight into the mechanism of primase function and are applicable for DNA primases fro

153 O'Brien et al proposed a novel mechanism of primase function based on redox activity of the iron-sul

154 The role of this region in primase function was further investigated by generating

155 We describe here a Trypanosoma brucei DNA primase gene, PRI1, that encodes a 70-kDa protein that l

156 Archaeal organisms contain conserved primase genes homologous to both the bacterial DnaG and

157 (ii) The T4 replisome with its primase (gp61) can also support priming and exponential

158 ls transfected with WT ICP8 and the helicase-primase (H/P) complex exhibited punctate nuclear structu

159 DNA primase harboring lysine at position 69 fails to stabili

160 et al report that the iron-sulfur cluster of primase has a redox role in enzyme activity.

161 as not been possible since a kinetoplast DNA primase has not been available.

162 and biochemical assays, we show that T7 DNA primase has only a slightly higher affinity for DNA cont

163 zinc-binding domain (ZBD) of prokaryotic DNA primases has been postulated to be crucial for recogniti

164 lthough structures of archaeal and bacterial primases have provided insights into general priming mec

165 ed sites were found within the UL5 (helicase-primase helicase subunit), UL23 (thymidine kinase), UL25

166 nal significance of their interactions using primase, helicase and primer extension assays, and a 'st

167 act with the beta-clamp), in the presence of primase, helicase, Pol III core, clamp loader, and beta-

168 ogy with the bacteriophage T7 gene 4 protein primase-helicase (T7 gp4).

169 Here we characterize a complex between T7 primase-helicase and DNA polymerase on DNA that was trap

170 Whereas one of the polymerases engages the primase-helicase and RNA primer on the lagging strand of

171 omplex consists of two DNA polymerases and a primase-helicase hexamer that assemble on the DNA templa

172 DNA polymerase and multiple subunits of the primase-helicase hexamer.

173 served within a bacterial class, whereas the primase-helicase interaction has co-evolved within each

174 The zinc binding domain of the primase-helicase is essential for trapping the RNA prime

175 These results indicate that the T7 primase-helicase specifically engages two copies of DNA

176 e long 52 gene (UL52; a component of the DNA primase/helicase complex), bICP4, IEtu2, and the unique

177 Pol-prim is composed of a primase heterodimer that synthesizes an RNA primer, a DN

178 Pol-prim consists of a primase heterodimer that synthesizes RNA primers, a DNA

179 quivalent peptide from human Pol alpha binds primase in an analogous fashion.

180 s that the DnaB-DnaC complex binds poorly to primase in comparison with DnaB alone.

181 , we describe the crystal structure of human primase in heterodimeric form consisting of full-length

182 well conserved with heterodimeric (p48/p58) primases in eukaryotes.

183 4-(2-pyridinyl)phenyl ]acetamide, a helicase-primase inhibitor for the treatment of herpes simplex vi

184 Pritelivir is a novel helicase-primase inhibitor in clinical development for treatment

185 ed novel herpes simplex virus (HSV) helicase-primase inhibitor that reduced genital shedding and lesi

186 Helicase-primase inhibitors are novel, potent inhibitors of herpe

187 Pol alpha/primase initiates primers on both strands that are exten

188 egulating primase activity, we characterized primase initiation specificity and interactions with the

189 ons that inhibit this charge transfer hinder primase initiation without affecting primase structure o

190 In eukaryotic and archaeal replication, primase is a heterodimer of two subunits, PriS and PriL.

191 with different mobilities only when helicase-primase is bound to DNA containing a single-stranded reg

192 harge transfer through the [4Fe4S] domain of primase is not feasible.

193 Bacterial primase is stimulated by replicative helicase to produce

194 single archaeo-eukaryotic primase (AEP), DNA primase, is required for the initiation and progression

195 of dATP, glycerol, and Tris buffer, the DNA primase isolated from Thermococcus kodakaraensis catalyz

196 n primosome and the C-terminal domain of the primase large subunit (p58C) with bound DNA/RNA duplex.

197 different proteins containing the helicase, primase, leading polymerase and a lagging strand polymer

198 The LTA primase LtaP(Lm) initiates LTA synthesis by transferring

199 The structures of p48 reveal that eukaryotic primases maintain the conserved catalytic prim fold doma

200 ults explain functional defects in human DNA primase mutants and provide insights into primosome load

201 Here we report the identification of a primase noncatalytic subunit, denoted PriX, from the hyp

202 A deficiency in the primase of bacteriophage T7 to synthesize primers can be

203 utagenesis of the zinc-binding domain of DNA primase of bacteriophage T7 using a bacterial homolog fr

204 ive tryptophan residues are dispersed in the primase of bacteriophage T7: Trp-42 in the ZBD and Trp-6

205 two of 12 potential priming sites of the DNA primase of the pRN1 replicon, but nearly all these mutat

206 h significant knowledge about single-subunit primases of prokaryotes has accumulated, the functions a

207 now be cited demonstrating how the term 'DNA primase' only describes a very narrow subset of these nu

208 port through DNA to the [4Fe4S] cluster of a primase p58C construct and a reversible switch in the DN

209 he N-terminal domain of the large subunit of primase (p58N) directly interacts with the C-terminal do

210 support of a functional bacterial-like DnaG primase participating in archaeal DNA replication, we ha

211 onstrated that the presence of a primer, not primase per se, provides the signal that triggers cyclin

212 DNA polymerase alpha-primase (pol-prim) plays a central role in DNA replicati

213 DNA polymerase alpha-primase (Pol-prim) plays an essential role in eukaryotic

214 plication protein A and DNA polymerase alpha-primase (pol-prim), constituting the viral primosome.

215 t low concentrations of DNA polymerase-alpha primase (Pol-primase), and the p58 subunit of Pol-primas

216 RPA-like ssDNA-binding complex, may regulate primase-Pol alpha (PP) activity at telomeres constitutiv

217 ochemical function of the CST complex is its primase-Pol alpha (PP) stimulatory activity.

218 a multifunctional replicative enzyme called primase-polymerase (PrimPol) that is capable of directly

219 we report that PrimPol, a recently described primase-polymerase (PrimPol), plays a crucial role in th

220 Here, we establish that Primase-Polymerase (PrimPol; CCDC111), an archaeal-eukar

221 PrimPol is a primase-polymerase found in humans, and other eukaryotes

222 ired for replication initiation, and the DNA primase-polymerase in eukaryotes is pol alpha.

223 PrimPol is a primase-polymerase involved in nuclear and mitochondrial

224 In many prokaryotes, a specific DNA primase/polymerase (PolDom) is required for nonhomologou

225 These structures, along with analysis of primase/polymerase activities, provide a plausible mecha

226 ion of these enzymes under a category called primase-polymerases within the wider functional grouping

227 ese findings establish that some replicative primases, previously considered to be solely involved in

228 Here, we report that archaeal replicative primases (Pri S, primase small subunit) can also perform

229 n enzymes: DNA polymerase delta (POLD1), DNA primase (PRIM1), and minichromosome 6 (MCM6).

230 kaging tegument protein), and UL52 (helicase-primase primase subunit) proteins.

231 ng activity of the bacteriophage T4 helicase-primase (primosome) complex.

232 bly, and function of the processive helicase-primase (primosome) component of the bacteriophage T4-co

233 g kinetics with those of the eukaryotic-type primase (PriSL) also found in Sso.

234 ms that guarantee DNA polymerase, clamp, and primase proteins are present for every cycle.

235 DNA primases provide oligoribonucleotides for DNA polymerase

236 ase), we developed the first non-radioactive primase-pyrophosphatase assay.

237 ghtly higher affinity for DNA containing the primase recognition sequence (5'-TGGTC-3') than for DNA

238 tations in T7 that suppress the inability of primase reduce the amount of gp5.5 and thus increase the

239 ural differences between bacterial and human primases render the former an excellent target for drug

240 st that the primase activity of the helicase-primase requires formation of a dimer or higher-order st

241 Alterations in subdomains of the primase result in loss of selective DNA binding.

242 Application of the model to DNA primase revealed a preference in the enzyme's second met

243 e crystal structure of the full-length human primase, revealing the precise overall organization of t

244 ision model) or synthesis of a new primer by primase (signaling model).

245 inding motif important in the recognition of primase sites in DNA.

246 t that archaeal replicative primases (Pri S, primase small subunit) can also perform TLS.

247 The addition of a single subunit of gp61 primase stabilized the resulting primosome complex at th

248 hinder primase initiation without affecting primase structure or polymerization.

249 ed inside the C-terminal domain of the large primase subunit (p58C).

250 and primase) activities when bound to a gp61 primase subunit at a helicase:primase subunit ratio of 6

251 nits of the helicase hexamer, and the single primase subunit interacts with both strands.

252 Recently, a third primase subunit named PriX was identified in the archaeo

253 ound to a gp61 primase subunit at a helicase:primase subunit ratio of 61.

254 A fifth SNV located within UL5 (helicase-primase subunit) greatly reduced in vivo viral replicati

255 rom six (gp41) helicase subunits, one (gp61) primase subunit, and nonhydrolyzable GTPgammaS.

256 s, together with a single tightly bound gp61 primase subunit.

257 The presence of additional primase subunits does not change the molecular mass or h

258 The mode of interaction of primase subunits with substrates during the various step

259 the large subunit of eukaryotic and archaeal primases, suggesting that the PhrB-like photolyases bran

260 Results demonstrated that primase template specificity is conserved within a bacte

261 l alpha, the critical interaction that keeps primase tethered to the eukaryotic replisome.

262 transcription factor (Mtf1) is an efficient primase that initiates DNA synthesis on ssDNA coated wit

263 synthesis in genomic duplication depends on primase, the DNA-dependent RNA polymerase that synthesiz

264 ulating the activity of DNA polymerase-alpha primase, the only enzyme known to initiate DNA replicati

265 DNA replication depends on primase, the specialised polymerase responsible for synt

266 d gp4 lacking the zinc binding domain of the primase; the protein has helicase activity but no DNA-de

267 Once loaded, the helicase recruits the primase through a direct protein-protein interaction to

268 fork construct prior to the addition of the primase to avoid the formation of metastable DNA-protein

269 The ability of T7 DNA primase to catalyze template-directed oligoribonucleotid

270 s an attractive candidate for serving as the primase to initiate lagging strand DNA synthesis during

271 in atomic detail the mode of association of primase to Pol alpha, the critical interaction that keep

272 imase, facilitating RNA primer handover from primase to Pol alpha.

273 and physiological significance of tethering primase to the eukaryotic replisome via pol alpha remain

274 /CTF1, suggesting that NFI also recruits Pol-primase to the NCCR.

275 These findings indicate that tethering of primase to the replisome by pol alpha is critical for th

276 s an oligoribonucleotide, synthesized by DNA primase, to initiate the synthesis of an Okazaki fragmen

277 been thought to require a protein, possibly primase, to pry polymerase from incompletely extended DN

278 tantially augments both primase activity and primase-to-polymerase switching.

279 domains of DNA helicase, five domains of RNA primase, two DNA polymerases, and two thioredoxin (proce

280 genome requires seven proteins: the helicase-primase (UL5-UL8-UL52), the DNA polymerase (UL30-UL42),

281 ngle-stranded DNA binding protein (UL29) and primase (UL52) genes.

282 icase-primase composed of helicase (UL5) and primase (UL52) subunits and a third subunit, UL8.

283 is revealed that upon binding Mn(II), T7 DNA primase undergoes conformational changes near the metal

284 Primases use single-stranded (ss) DNAs as templates to s

285 DnaB complexed with the C-terminal domain of primase, we found that Ile-85 is located at the interfac

286 ng NADH-quinone reductase subunit A, and DNA primase were expressed in HLA-B27(+) cells, and their HL

287 R screening, fragment molecules that bind T7 primase were identified and then exploited in virtual fi

288 Until relatively recently, DNA primases were viewed simply as a class of proteins that

289 at the RepA CTD interacts with the host DnaG primase, which binds the replicative helicase.

290 cantly enhances the binding of nucleotide to primase, which correlates with higher catalytic efficien

291 ures of the catalytic engine of a eukaryotic primase, which is contained in the p48 subunit.

292 ganisms, DNA replication is initiated by DNA primases, which synthesize primers that are elongated by

293 This direct interaction of a bacterial-like primase with a eukaryotic-like helicase suggests that fo

294 e, as well as the helicase-binding domain of primase with a molar ratio of 6:6:3 at 7.5 A resolution.

295 per DnaB hexamer and that the interaction of primase with DnaB and primer formation triggers the rele

296 The interaction of primase with DnaB and the release of DnaC mark discrete

297 peptide spanning the last 16 residues binds primase with high affinity, and the equivalent peptide f

298 has replaced PriL as the subunit that endows primase with the unique ability to initiate nucleic acid

299 The inclusion of multiple DNA primases within a whole domain of organisms complicates

300 Contributions of the primase zinc-binding domain, RNA polymerase domain and h