ライフサイエンスコーパス: MCM2

1 1) and minichromosome maintenance protein 2 (mcm2).

2 nished stimulation of DDK phosphorylation of Mcm2.

3 ective in stimulating DDK phosphorylation of Mcm2.

4 forks and interacts transiently with phospho-MCM2.

5 ators including E2F1, E2F2, EXO1, FOXM1, and MCM2.

6 saccharide fold (OB-fold) and A subdomain of Mcm2.

7 ells, with diminished DDK phosphorylation of Mcm2.

8 A family of six homologous subunits, Mcm2, -3, -4, -5, -6, and -7, each with its own unique f

9 e show that Cdt1 interacts with MCM subunits Mcm2, 4 and 6, which both destabilizes the Mcm2-5 interf

10 izes MCM in a left-handed spiral open at the Mcm2-5 gate.

11 s Mcm2, 4 and 6, which both destabilizes the Mcm2-5 interface and inhibits MCM ATPase activity.

12 The consequences of inappropriate Mcm2/5 gate actuation and the role of a side channel for

13 Recent cryo-EM structures of the Mcm2-7 (MCM) double hexamer, its precursors, and the ori

14 ing a prereplication complex that contains a Mcm2-7 (minichromosome maintenance proteins 2-7) double

15 the eukaryotic DNA replicative helicase, the Mcm2-7 (minichromosome maintenance) complex, is loaded a

16 e origin recognition complex (ORC)-Cdc6-Cdt1-Mcm2-7 (OCCM) intermediate showed that each of these sub

17 lysis promotes the formation of the ORC/Cdc6/MCM2-7 (OCM) complex, which functions in MCM2-7 double-h

18 ial' complex is transformed into an ORC/Cdc6/MCM2-7 (OCM) complex.

19 Although Mcm2-7 activation requires binding of the helicase-activ

20 activity was required for maximal loading of Mcm2-7 and a dramatic genome-wide reorganization of the

21 eplicate with decreased chromatin loading of MCM2-7 and become critically dependent on another ATPase

22 lication fork helicase is composed of Cdc45, Mcm2-7 and GINS (CMG).

23 replicative CMG helicase (comprising CDC45, MCM2-7 and GINS) occurs only after the final ligation st

24 ation as a ring-shaped heterohexamer to load MCM2-7 and initiate DNA replication.

25 0 stabilizes Cdc45 and GINS association with Mcm2-7 and stimulates replication elongation in vivo and

26 rom budding yeast, that Dpb11 alone binds to Mcm2-7 and that Dpb11 also competes with GINS for bindin

27 nd MSSB mutant combinations in S. cerevisiae Mcm2-7 are not viable.

28 re, origin melting and GINS association with Mcm2-7 are substantially decreased for cells expressing

29 red that inhibition of Cdc6 ATPase restricts MCM2-7 association with origin DNA to a single hexamer,

30 xes containing ATPase-motif mutations showed Mcm2-7 ATP binding and hydrolysis are required for helic

31 ic structure, our findings show that the six Mcm2-7 ATPase active sites are specialized for different

32 ic DNA replication is the recruitment of the MCM2-7 ATPase, the core of the replicative DNA helicase,

33 A subset of Mcm2-7 ATPase-site mutants completed helicase loading bu

34 Sld2-m1,4, that is specifically defective in Mcm2-7 binding.

35 that replication origins are not licensed by Mcm2-7 chromatin binding, but spindle disassembly occurs

36 e factors are overexpressed and effect clear Mcm2-7 chromatin binding.

37 hibitory activity, and consequently the Cdt1-MCM2-7 complex activates ORC/Cdc6 ATP-hydrolysis to prom

38 for intermediates consisting of an ORC-Cdc6-Mcm2-7 complex and an ORC-Cdc6-Mcm2-7-Mcm2-7 complex are

39 east, that Mcm10 directly interacts with the Mcm2-7 complex and Cdc45.

40 e complex protein Mcm4 alone and also of the Mcm2-7 complex and the dsDNA-loaded Mcm2-7 complex.

41 C-Cdc6-Mcm2-7 complex and an ORC-Cdc6-Mcm2-7-Mcm2-7 complex are reported, which together provide new

42 After Cdc45-Mcm2-7 complex assembly, Mcm10 promotes origin melting b

43 DNA as a single hexamer during ORC/Cdc6/Cdt1/MCM2-7 complex formation prior to MCM2-7 double hexamer

44 n origins prior to Cdt1 release and ORC-Cdc6-Mcm2-7 complex formation, but how the second Mcm2-7 hexa

45 ave other roles, the essential nature of the MCM2-7 complex in DNA replication has made it difficult

46 ulating the assembly and distribution of the Mcm2-7 complex in the Drosophila genome.

47 In fact, Mcm10 recruits Cdc45 to Mcm2-7 complex in vitro.

48 uitment of Cdc45 and GINS recruitment to the Mcm2-7 complex in vivo during early S phase.

49 ing consists of the loading of the hexameric MCM2-7 complex onto chromatin during G1 phase and is dep

50 complex, but not the 'initial' ORC/Cdc6/Cdt1/MCM2-7 complex, is competent for MCM2-7 dimerization.

51 eukaryotic replisome components include the Mcm2-7 complex, the CMG helicase, DNA polymerases, a Ctf

52 7 hexamer to form an 'initial' ORC/Cdc6/Cdt1/MCM2-7 complex.

53 o of the Mcm2-7 complex and the dsDNA-loaded Mcm2-7 complex.

54 nished GINS and pol-alpha recruitment to the Mcm2-7 complex.

55 tes are on predicted exposed surfaces of the MCM2-7 complex.

56 involved in the recruitment of Cdc45 to the Mcm2-7 complex.

57 pendent fashion with kinetics similar to the MCM2-7 complex.

58 th RNA polymerase induce a redistribution of Mcm2-7 complexes along the chromosomes, resulting in a c

59 tent with the current hypothesis that excess MCM2-7 complexes are loaded during G1 phase, and are req

60 Mutant Mcm2-7 complexes assemble and are recruited to replicati

61 Mcm2-7 complexes containing ATPase-motif mutations showe

62 These excess Mcm2-7 complexes exhibit little co-localization with ORC

63 mplexes (pre-RCs) license origins by loading Mcm2-7 complexes in inactive form around DNA.

64 n complex (ORC) assemble two heterohexameric Mcm2-7 complexes into a head-to-head double hexamer that

65 licase loading but instead drives removal of Mcm2-7 complexes that cannot complete loading.

66 e result of sequential loading of individual Mcm2-7 complexes.

67 ion are still debated, it interacts with the Mcm2-7 core helicase, the lagging strand polymerase, DNA

68 Here, we report the cryo-EM structure of the Mcm2-7 DH on dsDNA and show that the DNA is zigzagged in

69 To convert the Mcm2-7 DH structure into the Mcm2-7 hexamer structure fo

70 The MCM2-7 dimer, in contrast to the MCM2-7 double-hexamer,

71 s competent for MCM2-7 dimerization, reveals MCM2-7 dimerization as a limiting step during pre-RC for

72 identifies the OCM complex as competent for MCM2-7 dimerization, reveals MCM2-7 dimerization as a li

73 utants we discovered a complex competent for MCM2-7 dimerization.

74 C/Cdc6/Cdt1/MCM2-7 complex, is competent for MCM2-7 dimerization.

75 The regulated loading of the Mcm2-7 DNA helicase (comprising six related subunits, Mc

76 se, replisomes assemble around the activated Mcm2-7 DNA helicase.

77 cid alteration in MCM4 that destabilizes the MCM2-7 DNA replicative helicase, has fewer dormant repli

78 DDK phosphorylation of Mcm2-7 does not by itself promote separation of the doub

79 tin environment restricts the loading of the Mcm2-7 double hexamer either upstream of or downstream f

80 To determine whether Cdc6 regulates MCM2-7 double hexamer formation, we analysed complex ass

81 /Cdc6/Cdt1/MCM2-7 complex formation prior to MCM2-7 double hexamer formation.

82 but it is unknown how these proteins control MCM2-7 double hexamer formation.

83 w that the head-to-head juxtaposition of the Mcm2-7 double hexamer generates a new protein interactio

84 py, we report a near-atomic structure of the MCM2-7 double hexamer purified from yeast G1 chromatin.

85 uding RNA polymerase, can push budding yeast Mcm2-7 double hexamers along DNA.

86 eplication origins are "licensed" by loading MCM2-7 double hexamers and during S phase licensed repli

87 ric CMGs establishes the subunit register of Mcm2-7 double hexamers and together with the spiral form

88 These experiments indicate that Mcm2-7 double hexamers can be precursors of replication

89 dictate Mcm2-7 loading specificity and that Mcm2-7 double hexamers form preferentially at a native o

90 Displaced Mcm2-7 double hexamers support DNA replication initiatio

91 nize and encircle origin DNA and assemble an Mcm2-7 double-hexamer around adjacent double-stranded DN

92 RC) assembly after OCM formation, but before MCM2-7 double-hexamer assembly.

93 dc6/MCM2-7 (OCM) complex, which functions in MCM2-7 double-hexamer assembly.

94 Detailed structural analysis of the loaded Mcm2-7 double-hexamer complex demonstrates that the two

95 ly is inhibited, which is unexpected, as the MCM2-7 double-hexamer represents a very large interactio

96 The MCM2-7 dimer, in contrast to the MCM2-7 double-hexamer, interacts with ORC/Cdc6 and is sa

97 how the OCM is subsequently converted into a MCM2-7 double-hexamer.

98 eplicative complexes (pre-RCs), resulting in MCM2-7 double-hexamers on DNA.

99 ring S phase, impaired GINS interaction with Mcm2-7 during S phase, and decreased replication protein

100 s in increased interaction between Dpb11 and Mcm2-7 during S phase, impaired GINS interaction with Mc

101 kedly disrupted interaction between GINS and Mcm2-7 during S phase.

102 KEAP1 is thus poised to affect MCM2-7 dynamics or function rather than MCM3 abundance.

103 interaction could provide key insights into Mcm2-7 function and regulation.

104 The replicative helicase Mcm2-7 functions in both initiation and fork progression

105 The MCM2-7 genes are coordinately expressed during developme

106 Overexpression of MCM2-7 genes correlated with poor prognosis in breast ca

107 ntly correlated with an increasing number of MCM2-7 genes overexpression.

108 heterozygous loss or mutation of one or more MCM2-7 genes, which is expected to compromise DNA replic

109 inding experiments confirmed that CMG and/or Mcm2-7 had to be phosphorylated for binding to phospho-T

110 Furthermore, dsDNA-loaded Mcm2-7 harboring the DDK phosphomimetic Mcm4 mutant boun

111 Importantly, we demonstrate that the MCM2-7 helicase becomes loaded onto DNA as a single hexa

112 g G1-phase of the cell-cycle the replicative MCM2-7 helicase becomes loaded onto DNA into pre-replica

113 Cdc45) and GINS proteins activate the latent Mcm2-7 helicase by inducing allosteric changes through b

114 defined by the ORC-dependent loading of the Mcm2-7 helicase complex onto chromatin in G1.

115 target of the Dbf4/Cdc7 kinase (DDK) is the Mcm2-7 helicase complex.

116 d into two biochemically discrete steps: the Mcm2-7 helicase is first loaded into prereplicative comp

117 recognition complex (ORC), which coordinates Mcm2-7 helicase loading to form the prereplicative compl

118 oading, but also for the distribution of the Mcm2-7 helicase prior to S-phase entry.

119 Cdc45 activates the Mcm2-7 helicase.

120 This origin licensing requires loading two Mcm2-7 helicases around origin DNA in a head-to-head ori

121 s ORC-Cdc6 function to recruit a single Cdt1-Mcm2-7 heptamer to replication origins prior to Cdt1 rel

122 The Mcm2-7 heterohexamer, like other hexameric helicases, is

123 g yeast is the association of Cdc45 with the Mcm2-7 heterohexameric ATPase, and a second step is the

124 establish that the Saccharomyces cerevisiae MCM2-7 hexamer assumes a closed ring structure, suggesti

125 Although the MCM2-7 hexamer can adopt a ring shape with a gap between

126 t, show that ATPase activity is required for MCM2-7 hexamer dimerization and demonstrate that MCM2-7

127 During S phase, each Mcm2-7 hexamer forms the core of a replicative DNA helic

128 the active helicase, the N-tier ring of the Mcm2-7 hexamer in the DH-dsDNA needs to tilt and shift l

129 Mcm2-7 complex formation, but how the second Mcm2-7 hexamer is recruited to promote double-hexamer fo

130 To convert the Mcm2-7 DH structure into the Mcm2-7 hexamer structure found in the active helicase, t

131 Cdc6 recruits with the help of Cdt1 a single MCM2-7 hexamer to form an 'initial' ORC/Cdc6/Cdt1/MCM2-7

132 tive Cdc6 ATPase promotes recruitment of two MCM2-7 hexamer to origin DNA.

133 Through analysis of MCM2-7 hexamer-interface mutants we discovered a complex

134 tion, the core component of the helicase-the Mcm2-7 hexamer-is loaded on origin DNA as a double hexam

135 ents a specific conformational change in the Mcm2-7 hexamer.

136 dc45, Mcm2-7, GINS) helicase consists of the Mcm2-7 hexameric ring along with five accessory factors.

137 -7 hexamer dimerization and demonstrate that MCM2-7 hexamers are recruited to origins in a consecutiv

138 Here we show that both Mcm2-7 hexamers in Saccharomyces cerevisiae are recruite

139 n complex (ORC), Cdc6, and Cdt1 assemble two MCM2-7 hexamers into one double hexamer around dsDNA.

140 Finally, we demonstrate that single Mcm2-7 hexamers propagate bidirectionally, monotonically

141 Helicase loading involves two MCM2-7 hexamers that assemble into a double hexamer arou

142 These proteins load MCM2-7 in an unknown way into a double hexamer around DN

143 it, even its ATPase subunit, can load enough MCM2-7 in partnership with CDC6 to initiate DNA replicat

144 e BRCT4 motif of Dpb11 that remains bound to Mcm2-7 in the presence of ssDNA (dpb11-m1,m2,m3,m5), and

145 ucleotide ssDNA, and recruiting pol alpha to Mcm2-7 in vitro.

146 o, Ichi, Ni, and San (GINS) association with Mcm2-7 in vivo.

147 o test this model and assess the location of Mcm2-7 initial loading, we placed DNA-protein roadblocks

148 Thus, the origin melting and GINS-Mcm2-7 interaction defects we observed for mcm10-m2,3,4

149 and find that sld2-DNA cells exhibit no GINS-Mcm2-7 interaction.

150 ecognition complex (ORC), Cdc6 and Cdt1 load Mcm2-7 into a double hexamer bound around duplex DNA in

151 However, while eukaryotic MCM2-7 is a heterocomplex made of different polypeptide

152 MCM2-7 is activated by both the CDC7-Dbf4 kinase and cyc

153 ve helicase to the replication origin, where MCM2-7 is activated to initiate DNA replication.

154 These data suggest that Sld2 binding to Mcm2-7 is essential to block the inappropriate formation

155 RNase A sensitive, whereas interaction with Mcm2-7 is nuclease resistant.

156 The minimal amounts of Mcm2-7 loaded in the absence of cyclin E/Cdk2 activity w

157 In contrast, a Cdc6 sensor-1 mutant affects MCM2-7 loading and Cdt1 release, similar as a Cdc6 Walke

158 of Orc3 and is required for ORC function and MCM2-7 loading in vivo.

159 dynamics of the ORC-Cdc6 interaction dictate Mcm2-7 loading specificity and that Mcm2-7 double hexame

160 er the course of G1 is not only critical for Mcm2-7 loading, but also for the distribution of the Mcm

161 ATP hydrolysis can promote Mcm2-7 loading, but can also promote Mcm2-7 release if c

162 vity is known to facilitate Cdt1 release and MCM2-7 loading, we discovered that Orc1 ATP-hydrolysis i

163 Loading of each Mcm2-7 molecule involves the ordered association and dis

164 Loading-defective Mcm2-7 mutant complexes were defective in initial Mcm2-7

165 a detailed analysis of Arabidopsis thaliana mcm2-7 mutants and reveal phenotypic differences.

166 We demonstrate that these MCM2-7 mutants arrest during prereplicative complex (pre

167 ndependent, CDC6-dependent mechanism to load MCM2-7 on origins of replication.

168 al how ORC, Cdc6, and Cdt1 cooperate to load MCM2-7 onto DNA, enabling bidirectional replication.

169 or mutations that bypass the requirement for Mcm2-7 phosphorylation by DDK restored PFA in the absenc

170 es the polarity by which DNA enters into the Mcm2-7 pore, and explains how Cdc45 helps prevent DNA fr

171 Here we show that TIM interacts with MCM2-7 prior to the initiation of DNA replication.

172 Several aminoacyl-tRNA synthetases and Mcm2-7 proteins were identified by mass spectrometry of

173 Contrary to the sites of Mcm2-7 recruitment being precisely defined, only single

174 Here we show that MCM2-7 recruitment by ORC/Cdc6 is blocked by an autoinhi

175 7 mutant complexes were defective in initial Mcm2-7 recruitment or Cdt1 release.

176 rate how conserved Cdc6 AAA+ motifs modulate MCM2-7 recruitment, show that ATPase activity is require

177 ding, influence the ORC-Cdc6 interaction and MCM2-7 recruitment.

178 Paradoxically, there is a vast excess of Mcm2-7 relative to ORC assembled onto chromatin in G1.

179 s a 10-fold decrease in chromatin-associated Mcm2-7 relative to the levels found at the G1/S transiti

180 promote Mcm2-7 loading, but can also promote Mcm2-7 release if components are missing or if ORC has b

181 Activation of the Mcm2-7 replicative DNA helicase is the committed step in

182 tiate DNA replication by phosphorylating the Mcm2-7 replicative helicase [5-7].

183 Loading of the ring-shaped Mcm2-7 replicative helicase around DNA licenses eukaryot

184 ase active site (mcm2DENQ), we show that the Mcm2-7 replicative helicase has a novel DRC function as

185 The Mcm2-7 replicative helicase is central to all steps of e

186 plication licensing factor CDC6 recruits the MCM2-7 replicative helicase to the replication origin, w

187 single-molecule FRET, arrival of the second Mcm2-7 results in rapid double-hexamer formation that an

188 f4-Cdc7 phosphorylation of Mcm2 may open the Mcm2-7 ring at the Mcm2-Mcm5 interface, allowing for sin

189 between GINS/Cdc45 and the outer edge of the Mcm2-7 ring for unwinding have remained unexplored.

190 d Dbf4-Cdc7 phosphorylation of Mcm2 promotes Mcm2-7 ring opening.

191 ed in the interface between these domains in Mcm2-7 structures, mutations predicted to separate the d

192 The Mcm2-7 subcomplex forms a cracked-ring, right-handed spi

193 ically examined the interaction between each Mcm2-7 subunit with Dbf4 and Cdc7 through two-hybrid and

194 , it is not well understood which of the six Mcm2-7 subunits actually mediate(s) docking of this kina

195 his study demonstrates the importance of the MCM2-7 subunits during seed development and suggests tha

196 eukaryotes, the MSSB is conserved in several Mcm2-7 subunits, and MSSB mutant combinations in S. cere

197 e-wide reorganization of the distribution of Mcm2-7 that is shaped by active transcription.

198 together with the spiral form highlights how Mcm2-7 transitions through different conformational and

199 trusion of ssDNA from the central channel of Mcm2-7 triggers GINS attachment to Mcm2-7.

200 ld3/Treslin coordinates Cdc45 recruitment to Mcm2-7 with DDK phosphorylation of Mcm2 during S phase.

201 uble hexamers of minichromosome maintenance (Mcm2-7) at origin sites.

202 engage the mini-chromosome maintenance 2-7 (MCM2-7) complex during replicative helicase loading; how

203 The MINICHROMOSOME MAINTENANCE 2-7 (MCM2-7) complex, a ring-shaped heterohexamer, unwinds th

204 heterohexameric minichromosome maintenance (MCM2-7) helicase complex at replication origins during G

205 replicative mini-chromosome-maintenance 2-7 (MCM2-7) helicase is loaded in Saccharomyces cerevisiae a

206 ant loss of the mini-chromosome maintenance (Mcm2-7) helicase proteins on chromatin.

207 ase minichromosome maintenance proteins 2-7 (MCM2-7) onto replication origins is a prerequisite for r

208 We focus on the minichromosome maintenance (MCM2-7) proteins, which form the core of the eukaryotic

209 5 (Cdc45) to minichromosome maintenance 2-7 (Mcm2-7).

210 e 45 (Cdc45)/minichromosome maintenance 2-7 (Mcm2-7)/Go, Ichi, Nii, and San (GINS) (CMG) proteins [hu

211 al channel of Mcm2-7, Dpb11 dissociates from Mcm2-7, and Dpb11 binds to ssDNA, thereby allowing GINS

212 helicase in eukaryotes is composed of Cdc45, Mcm2-7, and GINS (CMG).

213 on fork helicase is comprised of CMG (Cdc45, Mcm2-7, and GINS) in eukaryotic cells, and the mechanism

214 MG replicative helicase (comprised of Cdc45, MCM2-7, and GINS), which obstructs the underlying cross-

215 helicase in eukaryotes is composed of Cdc45, Mcm2-7, and GINS.

216 Dpb11, although bound to Cdc45.Mcm2-7, can block the interaction between GINS and Mcm2-

217 trusion of ssDNA from the central channel of Mcm2-7, Dpb11 dissociates from Mcm2-7, and Dpb11 binds t

218 ve CMG [Cdc45 (cell division cycle gene 45), Mcm2-7, GINS (Go, Ichi, Ni, and San)] helicase.

219 The eukaryotic CMG (Cdc45, Mcm2-7, GINS) helicase consists of the Mcm2-7 hexameric

220 cation defect with no recruitment of GINS to Mcm2-7, whereas expression of wild-type levels of sld3-m

221 e-activation factors the assembly of a Cdc45-MCM2-7-GINS (CMG) complex.

222 blished recruitment mechanisms whereby Cdc45-Mcm2-7-GINS (CMG) helicase binds Pol epsilon and tethers

223 Assembly of the Cdc45-Mcm2-7-GINS (CMG) replicative helicase complex must be r

224 ion are the assembly and activation of Cdc45-Mcm2-7-GINS (CMG) replicative helicase.

225 f an ORC-Cdc6-Mcm2-7 complex and an ORC-Cdc6-Mcm2-7-Mcm2-7 complex are reported, which together provi

226 sDNA, thereby allowing GINS to bind to Cdc45.Mcm2-7.

227 model in which Dpb11 first recruits Cdc45 to Mcm2-7.

228 the loading of the replicative DNA helicase, Mcm2-7.

229 , can block the interaction between GINS and Mcm2-7.

230 Dpb11 also competes with GINS for binding to Mcm2-7.

231 ished Go, Ichi, Ni, and San association with Mcm2-7.

232 e also found that Dpb11 can recruit Cdc45 to Mcm2-7.

233 hannel of Mcm2-7 triggers GINS attachment to Mcm2-7.

234 extrusion and subsequent GINS assembly with Mcm2-7.

235 y disrupts the interaction between Cdc45 and Mcm2-7.

236 anism to recruit Dpb11 to DDK-phosphorylated Mcm2-7.

237 case loading or in releasing ORC from loaded MCM2-7.

238 trameric GINS (GG-Ichi-Nii-San) complex with Mcm2-7.

239 2, which thereby leads to GINS attachment to Mcm2-7.

240 ssembly, and the recruitment of pol alpha to Mcm2-7.

241 ing a mechanism for how GINS is recruited to Mcm2-7.

242 lication defect with no Cdc45 recruitment to Mcm2-7.

243 nd ssDNA inhibits the Dpb11 interaction with Mcm2-7.

244 ichi-ni-san complex (GINS) to form the CDC45.MCM2-7.GINS (CMG) helicase complex.

245 ric changes through binding, forming a Cdc45/Mcm2-7/GINS (CMG) complex that is competent to unwind du

246 The Cdc45/Mcm2-7/GINS (CMG) helicase separates DNA strands during

247 s but do not associate with the active CDC45/MCM2-7/GINS (CMG) replicative helicase.

248 n origin activation by regulating CMG (CDC45/MCM2-7/GINS) helicase.

249 -RC) assembly by loading a double hexamer of Mcm2-7: the core of the replicative helicase.

250 rylation of the replicative helicase subunit MCM2, an ATR effector.

251 treatment, along with loss of expression of MCM2 and CDK1, and reduction in dNTP levels.

252 of cell cycle-associated proteins including MCM2 and cyclins A, E, D1/D3 in macrophages, without evi

253 dc7 phosphorylates the known Cdc7 substrates Mcm2 and histone H3 in vitro, and Cdc7 kinase activity i

254 The immobile fraction of MCM2 and MCM4 increases during G1 phase, suggestive of r

255 We show that, in telophase, MCM2 and MCM4 maintain transient interactions with chrom

256 lted in synthetic lethality, suggesting that Mcm2 and Mcm4 play overlapping roles in the association

257 ugh a unique DNA entry gate comprised of the Mcm2 and Mcm5 subunits.

258 The CMG has an active gate between subunits Mcm2 and Mcm5 that opens and closes in response to nucle

259 ubstantially weakens the interaction between Mcm2 and Mcm5, and Dbf4-Cdc7 phosphorylation of Mcm2 pro

260 er can adopt a ring shape with a gap between Mcm2 and Mcm5, it is unknown which Mcm interface functio

261 anked by two pairs of gate-forming subunits, MCM2 and MCM5.

262 e binding site for Mcm10 on MCM includes the Mcm2 and Mcm6 subunits and overlaps that for the loading

263 DNA at a faster rate compared to 9 degrees N MCM2 and to Mth MCM.

264 cation, reduced levels of DDK-phosphorylated Mcm2, and diminished Go, Ichi, Ni, and San (GINS) associ

265 ctive for stimulating DDK phosphorylation of Mcm2, binding to eighty-nucleotide ssDNA, and recruiting

266 Additionally, structures of Mcm2 bound to an H3/H4 tetramer suggest a direct role of

267 that Sld3 stimulates DDK phosphorylation of Mcm2 by 11-fold.

268 timulates human DDK phosphorylation of human Mcm2 by 15-fold.

269 nt does not stimulate the phosphorylation of Mcm2 by Dbf4-dependent kinase (DDK) in vitro.

270 Inhibiting Dbf4-Cdc7 phosphorylation of Mcm2 confers a dominant-negative phenotype with a severe

271 DDK phosphorylation of human Mcm2 decreases the affinity of Mcm5 for Mcm2, suggesting

272 ent strand capture and release and show that MCM2 deficiency reduces DNA replication initiation in ge

273 ocations that are preferentially affected by MCM2 deficiency.

274 of recurrent focal CNVs in tumors arising in MCM2-deficient mice, consistent with a direct relationsh

275 itment to Mcm2-7 with DDK phosphorylation of Mcm2 during S phase.

276 phosphorylates minichromosome maintenance 2 (Mcm2) during S phase in yeast, and Sld3 recruits cell di

277 e growth defect conferred by DDK-phosphodead Mcm2 in budding yeast.

278 The Dbf4-Cdc7 kinase phosphorylates Mcm2 in vitro, but the in vivo role for Dbf4-Cdc7 phosph

279 that budding yeast Dbf4-Cdc7 phosphorylates Mcm2 in vivo under normal conditions during S phase.

280 n vivo role for Dbf4-Cdc7 phosphorylation of Mcm2 is unclear.

281 Thus, Dbf4-Cdc7 phosphorylation of Mcm2 may open the Mcm2-7 ring at the Mcm2-Mcm5 interface

282 positions the DNA right in front of the two Mcm2-Mcm5 gates, with each strand being pressed against

283 tion of Mcm2 may open the Mcm2-7 ring at the Mcm2-Mcm5 interface, allowing for single-stranded DNA ex

284 tro, while in vivo analysis establishes that Mcm2/Mcm5 gate opening is essential for both helicase lo

285 Cells expressing either an Mcm2 mutant lacking this docking domain (Mcm2DeltaDDD) o

286 oxic sensitivity was found to be specific to Mcm2 or Mcm4 overexpression, further pointing to the imp

287 uld be induced in Mcm4DeltaDDD cells through Mcm2 overexpression as a means of titrating the Dbf4-MCM

288 ax), high MPP, and high blood flow expressed mcm2 (P = 0.036).

289 We also found that Mcm10 stimulates Mcm2 phosphorylation by DDK in vivo and in vitro.

290 mcm10-m2,3,4 are not explained by decreased Mcm2 phosphorylation by DDK, since the defects persist i

291 ls expressing sld3-m16 exhibit no detectable Mcm2 phosphorylation in vivo, reduced replication protei

292 We show here DDK-phosphoryled Mcm2 preferentially interacts with Cdc45 in vivo, and th

293 2 and Mcm5, and Dbf4-Cdc7 phosphorylation of Mcm2 promotes Mcm2-7 ring opening.

294 We identified an N-terminal Mcm2 region required for interaction with Dbf4.

295 o, Dbf4-Cdc7 kinase (DDK) phosphorylation of Mcm2 substantially weakens the interaction between Mcm2

296 uman Mcm2 decreases the affinity of Mcm5 for Mcm2, suggesting a potential mechanism for helicase ring

297 A helicase (comprising six related subunits, Mcm2 to Mcm7) into pre-replicative complexes at multiple

298 Inhibiting Dbf4-Cdc7 phosphorylation of Mcm2 under wild-type expression conditions also results

299 erved, as Dbf4 interacted most strongly with Mcm2, whereas Cdc7 displayed association with both Mcm4

300 elting by stimulating DDK phosphorylation of Mcm2, which thereby leads to GINS attachment to Mcm2-7.