ライフサイエンスコーパス: MCAK

1 MCAK (mitotic centromere-associated kinesin) is a Kin I

2 MCAK association with chromosome arms is promoted by pho

3 MCAK belongs to the Kin I subfamily of kinesin-related p

4 MCAK belongs to the Kinesin-13 family, whose members dep

5 MCAK depletion promoted dramatic spindle rocking in earl

6 MCAK has one high-affinity binding site per protofilamen

7 MCAK is a cognate substrate of PAK1.

8 MCAK is a homodimer that is encoded by a single gene and

9 MCAK is a member of the kinesin-13 family of microtubule

10 MCAK is a member of the kinesin-13 family of microtubule

11 MCAK localization and activity are regulated by Aurora B

12 MCAK microtubule depolymerization activity is inhibited

13 MCAK overexpression induces centromere-independent bundl

14 MCAK phosphorylation also regulates MCAK localization: t

15 MCAK targets protofilament ends very rapidly (on-rate 54

16 MCAK tracks with microtubule tips by binding to end-bind

17 MCAK, a Kinesin-13, catalytically depolymerizes microtub

18 ell-described catastrophe factors kinesin-13 MCAK and kinesin-8 Kip3/KIF18A.

19 n cells, contain the prototypical Kinesin-13 MCAK as well as a second family member, XKIF2.

20 The kinesin-13 MCAK is a potent MT depolymerase with a complex subcellu

21 which are governed in part by the kinesin-13 MCAK.

22 nce of the kinesin-8 Kip3 and the kinesin-13 MCAK.

23 We propose that up to 14 MCAK dimers assemble at the end of a microtubule to form

24 We propose a model in which KLP-7/MCAK regulates k-MT attachment and spindle tension to pr

25 a CENP-E mediated wall-tethering event and a MCAK-mediated wall-removing event, we establish that hum

26 Thus, a Rac1-Aurora A-MCAK signaling pathway mediates EC polarization and dire

27 hexylene glycol, but was unaffected by alpha-MCAK antibody and AMPPNP, which block catastrophe and ki

28 We find that although MCAK and XKIF2 have similar localization and biochemical

29 Prior to anaphase, ICIS localized in an MCAK-dependent manner to inner centromeres, the chromoso

30 microtubule formation at kinetochores in an MCAK-dependent manner.

31 d ), the kinesin-related proteins CENP-E and MCAK and the proposed structural and checkpoint proteins

32 kinase Aurora B also interacts with ICIS and MCAK raising the possibility that Aurora B may regulate

33 Kif2a and MCAK have documented roles in mitosis, but the function

34 epleted of the kinesin-13 proteins Kif2a and MCAK lack detectable flux and that such cells frequently

35 kinesin-13 proteins called Kif2a, Kif2b, and MCAK (Kif2c).

36 depolymerase activities of Kif2a, Kif2b, and MCAK fulfill distinct functions during mitosis in human

37 d Kif2a, play distinct roles in mitosis, and MCAK activity at kinetochores must be balanced by Kif2a

38 monopolar spindles, indicating that TOGp and MCAK contribute to spindle bipolarity, without major eff

39 Mitotic cells lacking both TOGp and MCAK formed bipolar and monopolar spindles, indicating t

40 interacts with microtubules and antagonizes MCAK activity, thus promoting bipolar spindle assembly.

41 eals that depletion of centromere-associated MCAK considerably decreases the directional coordination

42 nts designed to target centromere-associated MCAK for mechanistic analysis.

43 ENP and TD60, whereas a central region binds MCAK, Kif2a, and microtubules, suggesting a scaffold fun

44 luorescence localization of centromere-bound MCAK and found that MCAK localized to inner kinetochores

45 ochore microtubules from depolymerization by MCAK.

46 rturbation of spindle microtubule subsets by MCAK inhibition.

47 At inner centromeres, MCAK-ICIS may destabilize these microtubules and provide

48 Depletion of centromeric MCAK led to reduced centromere stretch, delayed chromoso

49 Codepleting MCAK with ch-Tog improved kinetochore fiber length and i

50 Xenopus extracts, ICIS coimmunoprecipitated MCAK and the inner centromere proteins INCENP and Aurora

51 In contrast, MCAK depletion rescued the proliferation of mutant pacli

52 In contrast, MCAK eliminated the aging process.

53 In contrast, MCAK inhibition caused a dramatic reorganization of spin

54 critical to temporally and spatially control MCAK function.

55 ive because of interference from cytoplasmic MCAK's global regulation of MT dynamics.

56 ct link between the microtubule depolymerase MCAK and Aurora B kinase.

57 tic activity of the microtubule depolymerase MCAK.

58 The MT depolymerase MCAK (mitotic centromere-associated kinesin) can influen

59 ), which stimulates the related depolymerase MCAK, can reactivate Kif2a after Aurora B inhibition.

60 walls and (2) the microtubule depolymerizer MCAK to release laterally attached microtubules after a

61 We show that the potent MT depolymerizer MCAK tracks (treadmills) with the tips of polymerizing M

62 ts were assayed to address how the different MCAK domains contribute to in vitro microtubule depolyme

63 or MCAK using FRAP analysis of the different MCAK mutants.

64 Aurora B biorients chromosomes by directing MCAK to depolymerize incorrectly oriented kinetochore mi

65 y chain, chromokinesin KIF4A, KIF3C, CENP-E, MCAK, and KIFC3) were not significantly inhibited by mil

66 se effects are reversed by anchoring ectopic MCAK to the centromere.

67 Expression of either MCAK (S/E) or MCAK (S/A) mutants increased the frequency

68 Substituting endogenous MCAK in Xenopus extracts with the alanine mutant XMCAK-4

69 Aurora B activity enriches MCAK at merotelic attachments and phosphorylates MCAK on

70 a result of MT instability caused by excess MCAK activity.

71 ole for Aurora B, which is to prevent excess MCAK binding to chromatin to facilitate chromatin-nuclea

72 In Xenopus extracts, MCAK associates with and is stimulated by the inner cent

73 e energy transfer (FRET)-based biosensor for MCAK and show that MCAK in solution exists in a closed c

74 Indeed, we found that hSgo2 is essential for MCAK to localize to the centromere.

75 Tip tracking is not essential for MCAK's MT-depolymerizing activity.

76 (.)tubulin complex formation as observed for MCAK.

77 first study that clearly defines a role for MCAK at the spindle poles as well as identifies another

78 lso detected two different binding sites for MCAK using FRAP analysis of the different MCAK mutants.

79 assembly and, further, cannot substitute for MCAK.

80 tudies revealed that the smallest functional MCAK deletion constructs are monomers.

81 sion using siRNA impaired the ability of GFP-MCAK to localize to MT tips in transfected cells.

82 a complex subcellular localization, yet how MCAK spatial regulation contributes to spindle assembly

83 However, MCAK's function at the centromere has remained mechanist

84 Here, we confirm that human MCAK colocalizes with EB1 at growing MT ends when expres

85 now show that the Kin I MCAK is a microtubule end-stimulated ATPase that can cat

86 IM and TIRF imaging, we find that changes in MCAK conformation are associated with a decrease in MCAK

87 nformation are associated with a decrease in MCAK affinity for the microtubule.

88 ins, we show that mitotic cells deficient in MCAK fail to maintain spindle bipolarity in the absence

89 rminal localization and regulatory domain in MCAK but not with the motor domain of the protein.

90 Collapse of bipolar spindles in MCAK-deficient cells is driven by pole-focusing activiti

91 ns involved in chromosome movement including MCAK, chromokinesin, and CENP-E may be descended from a

92 E715A/E716A in the far C-terminus increased MCAK targeting to the poles and reduced MT lifetimes, wh

93 Aurora B inhibited MCAK's microtubule depolymerizing activity in vitro, and

94 es MT stability during mitosis by inhibiting MCAK MT depolymerase activity.

95 tion of excess XMAP215 or EB1, or inhibiting MCAK (a Kinesin-13).

96 monstrate that this domain of Nup98 inhibits MCAK depolymerization activity in vitro.

97 Interestingly, MCAK accumulated at leading kinetochores during congress

98 kinesin-13 isoforms (Kif2a, Kif2b, and Kif2c/MCAK), which are highly conserved in their primary seque

99 y) are crucial for spindle formation; KifC1, MCAK (a member of the kinesin-13 family), CENP-E (a memb

100 MT stabilizer and the depolymerizing kinesin MCAK are differentially required for MT dynamics in the

101 The MT depolymerizing kinesin MCAK has also been reported to track growing MT plus end

102 on of the microtubule-depolymerizing kinesin MCAK, whose activity is negatively regulated by Aurora B

103 The KinI kinesin MCAK is a microtubule depolymerase important for governi

104 The KinI kinesin MCAK, a microtubule depolymerase, is critical for this r

105 inish centromere localization of the kinesin MCAK and the mitotic checkpoint response to taxol.

106 actor mitotic centromere-associated kinesin (MCAK) (a kinesin 13, previously called XKCM) and destabi

107 a and mitotic centromere-associated kinesin (MCAK) and inhibits their depolymerase activities.

108 erase mitotic-centromere-associated kinesin (MCAK) are required to release improper microtubule attac

109 sins, mitotic centromere-associated kinesin (MCAK) does not translocate along the surface of microtub

110 rizer mitotic centromere-associated kinesin (MCAK) from HeLa cells to produce ultra-long, astral MTs

111 in-13 mitotic centromere-associated kinesin (MCAK) increases chromosome misalignment and missegregati

112 erase mitotic centromere-associated kinesin (MCAK) is a key regulator for an accurate kinetochore-mic

113 Mitotic centromere-associated kinesin (MCAK) is a MAP that promotes MT disassembly within the m

114 Mitotic centromere-associated kinesin (MCAK) is a microtubule depolymerizer that is consistent

115 Mitotic centromere-associated kinesin (MCAK) is a microtubule-depolymerizing kinesin-13 member

116 Mitotic centromere-associated kinesin (MCAK) is recruited to the centromere at prophase and rem

117 osis, mitotic centromere-associated kinesin (MCAK) localizes to chromatin/kinetochores, a cytoplasmic

118 otein mitotic centromere-associated kinesin (MCAK) remains unknown.

119 that mitotic centromere-associated kinesin (MCAK), a kinesin-related protein that destabilizes micro

120 ty of mitotic centromere-associated kinesin (MCAK), thereby promoting leading-edge MT growth and cell

121 d the mitotic centromere-associated kinesin (MCAK).

122 izing mitotic centromere-associated kinesin (MCAK).

123 Mitotic centromere-associated kinesin (MCAK)/Kif2C is the most potent microtubule (MT)-destabil

124 ingle mitotic centromere-associated kinesin (MCAK)/kinesin-13 in Caenorhabditis elegans, is required

125 ells, mitotic centromere associated kinesin (MCAK; KIF2C) prevents chromosome segregation errors by d

126 Full-length MCAK exhibits higher ATPase activity, more efficient mic

127 or regions within the context of full-length MCAK is unknown.

128 ssays to compare the activity of full-length MCAK, which is a dimer, with MD-MCAK, which is a monomer

129 hromosomal passenger complex regulates local MCAK activity to permit spindle formation via stabilizat

130 Aurora B activity was required to localize MCAK to centromeres, but not to spindle poles.

131 ver, how the cytoplasmic- and pole-localized MCAK are regulated is currently not clear.

132 ytic domain plus the class specific neck (MD-MCAK), which is consistent with previous reports.

133 full-length MCAK, which is a dimer, with MD-MCAK, which is a monomer.

134 ule depolymerization assays, and microtubule.MCAK cosedimentation assays to compare the activity of f

135 Moreover, MCAK-depleted oocytes can recover from mono-orientation

136 Despite its apparent lack of motility, MCAK also contains a neck domain.

137 Thus, two KinI motors, MCAK and Kif2a, play distinct roles in mitosis, and MCAK

138 lymerizing activity observed in the neckless MCAK mutant.

139 o microtubule ends, enhancing the ability of MCAK to recycle for multiple rounds of microtubule depol

140 crotubule attachment may influence access of MCAK to Aurora B kinase and its opposing phosphatases.

141 essing during mitosis caused accumulation of MCAK, a microtubule depolymerase, on the spindle, indica

142 e notion that the antagonistic activities of MCAK and ch-Tog determine overall microtubule stability

143 sms must exist that modulate the activity of MCAK, both spatially and temporally.

144 Alteration of MCAK conformation by the point mutation E715A/E716A in t

145 tion of T95 on MCAK increases the binding of MCAK to centromeres.

146 Here, we show that the binding of MCAK to chromosome arms is also regulated by Aurora B an

147 We tested the consequences of MCAK disruption by injecting a centromere dominant-negat

148 eficient cells by simultaneous deficiency of MCAK or Nuf2 or treatment with low doses of nocodazole.

149 Delocalization of MCAK accounts for why cells depleted of hSgo2 exhibit ki

150 Depletion of MCAK altered mitotic spindle morphology, increased the f

151 Antisense-induced depletion of MCAK results in the same defect.

152 Depletion of MCAK, a kin I kinesin, increased MT lengths and density

153 Depletion of MCAK/Kif2C by siRNA stably decreases MT assembly rates i

154 Furthermore, we found that disruption of MCAK leads to multiple kinetochore-microtubule attachmen

155 e we show that the conserved motor domain of MCAK is necessary but not sufficient for microtubule dep

156 olyacrylamide ECMs to examine the effects of MCAK expression on MT growth dynamics and EC branching m

157 cs, its role in controlling the functions of MCAK remains unknown.

158 rotubule assembly, a function independent of MCAK activity.

159 le-focusing activities and is independent of MCAK function at centromeres, implicating hyperstabilize

160 Thus, GTSE1 inhibition of MCAK activity regulates the balance of MT stability that

161 pindle poles and an impaired localization of MCAK and HURP, two key regulators of mitotic spindle for

162 h the correction of mitotic defects, loss of MCAK reversed an aberrantly high frequency of microtubul

163 crotubule plus ends at kinetochores (loss of MCAK).

164 in vitro, and phospho-mimic (S/E) mutants of MCAK inhibited depolymerization in vivo.

165 ea that multiple phosphorylation pathways of MCAK cooperate to spatially control MT dynamics to maint

166 urora B, but how Aurora B phosphorylation of MCAK affects spindle assembly is unclear.

167 rmore, we found that PAK1 phosphorylation of MCAK on serines 192 and 111 preferentially regulates its

168 the microtubule tip-associated population of MCAK: negative regulation of microtubule length within t

169 so play an important role in processivity of MCAK-induced MT depolymerization.

170 ndent of PP2A and mediated by recruitment of MCAK and inhibition of Aurora C kinase activity respecti

171 d is only modestly sensitive to reduction of MCAK action.

172 rylation of serine 196 in the neck region of MCAK inhibited its microtubule depolymerization activity

173 ving an Aurora B-PLK1 axis for regulation of MCAK activity in mitosis.

174 it contributes to the spatial regulation of MCAK activity within inner centromere and kinetochore.

175 ate turnover is crucial in the regulation of MCAK activity.

176 Here, we report the regulation of MCAK by Aurora B.

177 Regulation of MCAK function is dependent on Aurora A kinase, which is

178 mechanistic insight into PAK1 regulation of MCAK functions.

179 rapid rapamycin-dependent relocalization of MCAK/Kif2C and Kif18A to the plasma membrane.

180 This localization pattern is reminiscent of MCAK, which is a microtubule depolymerase that is believ

181 onal image analysis to elucidate the role of MCAK in regulating MT growth dynamics, morphology, and d

182 changed, suggesting that the primary role of MCAK is not to move chromosomes.

183 Here, we compare the roles of MCAK and XKIF2 during spindle assembly in Xenopus extrac

184 The functional significance of MCAK's tip-tracking behavior during mitosis has never be

185 ay between multiple phosphorylation sites of MCAK may be critical to temporally and spatially control

186 Here, we determine the structure of MCAK motor domain bound to its regulatory C-terminus.

187 Furthermore, substitution of MCAK's neck domain with either the positively charged KI

188 pendent on the extreme COOH-terminal tail of MCAK.

189 our work shows how the regional targeting of MCAK regulates MT dynamics, highlighting the idea that m

190 Although targeting of MCAK to centromeres requires phosphorylation of S110 on

191 t or RNAi prevented centromeric targeting of MCAK.

192 Here we show that the far C-terminus of MCAK plays a critical role in regulating MCAK conformati

193 Tip tracking of MCAK is inhibited by phosphorylation and is dependent on

194 minus to permit diffusional translocation of MCAK along the surface of MTs.

195 We defined the minimal functional unit of MCAK as the catalytic domain plus the class specific nec

196 A motorless version of MCAK that binds centromeres but not microtubules disrupt

197 at this depolymerization is not dependent on MCAK dimerization.

198 This function depends on MCAK's ability to bind EBs and track with polymerizing n

199 tromeres requires phosphorylation of S110 on MCAK, dephosphorylation of T95 on MCAK increases the bin

200 on MCAK, whereas phosphorylation of S196 on MCAK promotes dissociation from the arms.

201 mapped six Aurora B phosphorylation sites on MCAK in both the centromere-targeting domain and the nec

202 of S110 on MCAK, dephosphorylation of T95 on MCAK increases the binding of MCAK to centromeres.

203 rms is promoted by phosphorylation of T95 on MCAK, whereas phosphorylation of S196 on MCAK promotes d

204 B phosphorylation at S196 in the neck opens MCAK conformation and diminishes the interaction between

205 Expression of either MCAK (S/E) or MCAK (S/A) mutants increased the frequency of syntelic m

206 K1 signaling at the kinetochore orchestrates MCAK activity, which is essential for timely correction

207 vity or expression of a non-phosphorylatable MCAK mutant prevents correct kinetochore-microtubule att

208 at merotelic attachments and phosphorylates MCAK on residues that regulate its microtubule depolymer

209 PAK1 phosphorylates MCAK and thereby regulates both its localization and fun

210 ubstrate of PAK1 wherein PAK1 phosphorylates MCAK on serines 192 and 111 both in vivo and in vitro.

211 Active PLK1, in turn, phosphorylates MCAK at Ser715 which promotes its microtubule depolymera

212 the regulatory mechanism underlying precise MCAK depolymerase activity control during mitosis remain

213 In this study, we present MCAK chimeras and mutants designed to target centromere-

214 microtubule depolymerization, and preventing MCAK from being sequestered by tubulin heterodimers.

215 portant for its catalytic cycle by promoting MCAK binding to microtubule ends, enhancing the ability

216 of KLP-7 or the mammalian kinesin-13 protein MCAK (KIF2C) also resulted in ectopic microtubule asters

217 ressor protein APC or the kinesin-13 protein MCAK, is sufficient to promote chromosome segregation de

218 The kinesin-13 motor protein MCAK is a potent microtubule depolymerase.

219 Rather, MCAK, but not XKIF2, plays a central role in regulating

220 g the possibility that Aurora B may regulate MCAK activity as well.

221 MCAK phosphorylation also regulates MCAK localization: the MCAK (S/E) mutant frequently loca

222 w that Aurora B phosphorylates and regulates MCAK both in vitro and in vivo.

223 Aurora B regulates MCAK's activity and localization.

224 a B both positively and negatively regulates MCAK during mitosis.

225 y to MCAK function, with Aurora B regulating MCAK's activity and its localization at the centromere a

226 of MCAK plays a critical role in regulating MCAK conformation, subspindle localization, and spindle

227 th rapid and long-term loss of MT regulators MCAK/Kif2C and Kif18A.

228 Unexpectedly, robust MCAK microtubule (MT) depolymerization activity is not n

229 e identified ICIS, a protein that stimulates MCAK activity in vitro.

230 Phosphorylation switches MCAK conformation, which inhibits its ability to interac

231 Purified GFP-tagged MCAK domain mutants were assayed to address how the diff

232 d chromosome arms in mid-meiosis I, and that MCAK depletion, or inhibition using a dominant-negative

233 These results demonstrate that MCAK undergoes long-range conformational changes involvi

234 is targeted to growing MT ends by EB1, that MCAK is held in an inactive conformation when associated

235 We find that MCAK is recruited to centromeres, kinetochores and chrom

236 In particular, we found that MCAK colocalized with NuMA and XMAP215 at the center of

237 Here we found that MCAK is a cognate substrate of PAK1 wherein PAK1 phospho

238 of chromatin and centrosomes and found that MCAK localization and activity are tightly regulated by

239 In addition, we found that MCAK localization at spindle poles was regulated through

240 tion of centromere-bound MCAK and found that MCAK localized to inner kinetochores during prophase but

241 Here, we tested the hypothesis that MCAK contributes to compliance and dimensionality mechan

242 Our results identify that MCAK promotes fast MT growth speeds in ECs cultured on c

243 The results indicate that MCAK affects cell sensitivity to mitotic inhibitors by m

244 Live cell imaging indicates that MCAK may be required to coordinate the onset of sister c

245 Our data support the model that MCAK perturbs spindle organization by acting preferentia

246 We propose that MCAK activity is spatially controlled by an interplay be

247 We propose that MCAK increases the turnover of kinetochore MTs at all ce

248 We propose that MCAK is targeted to growing MT ends by EB1, that MCAK is

249 We show that MCAK associates with the C-terminus of EB1 and EB3 but m

250 escence lifetime imaging (FLIM) we show that MCAK bound to microtubule ends is closed relative to MCA

251 FRET)-based biosensor for MCAK and show that MCAK in solution exists in a closed conformation mediate

252 We show that MCAK is an ATPase that catalytically depolymerizes micro

253 Our results show that MCAK regulation of cytoplasmic and spindle-associated mi

254 Our results show that MCAK-mediated depolymerization of MTs is specifically ta

255 Our studies suggest that MCAK dimerization is important for its catalytic cycle b

256 otubule catastrophe like that induced by the MCAK Kinesin-13s.

257 represents a structural intermediate in the MCAK catalytic cycle.

258 conserved positively charged residues in the MCAK neck domain significantly reduced MT depolymerizati

259 lts also indicate that plant kinesins in the MCAK/Kinesin-13 subfamily have evolved to take on differ

260 otor kinesins of animals and protists in the MCAK/Kinesin13 subfamily.

261 lation also regulates MCAK localization: the MCAK (S/E) mutant frequently localized to the inner cent

262 Our analysis reveals that the MCAK C-terminus binds to two motor domains in solution a

263 Thus, MCAK contributes to chromosome alignment in meiosis I, b

264 These results link Aurora B activity to MCAK function, with Aurora B regulating MCAK's activity

265 nd to microtubule ends is closed relative to MCAK associated with the microtubule lattice.

266 t spindle length is exquisitely sensitive to MCAK concentration but not XKIF2 concentration.

267 zation signal SKIP, which lies N terminal to MCAK's neck and motor domain.

268 re, we find that 3D ECM engagement uncouples MCAK-mediated regulation of MT growth persistence from m

269 of both phosphorylated and unphosphorylated MCAK protein, suggesting that phosphate turnover is cruc

270 aximum lengths prior to catastrophe, whereas MCAK promotes rapid restructuring of the microtubule cyt

271 ction in mouse oocyte meiosis I, and whether MCAK is necessary to prevent chromosome segregation erro

272 Here, we examine whether MCAK is involved in spindle function in mouse oocyte mei

273 se that tip tracking is a mechanism by which MCAK is preferentially localized to regions of the cell

274 We set out to understand why MCAK and Aurora B are more abundant at some metaphase-al

275 ults define a highly conserved domain within MCAK and related (KIN I) kinesins that is critical for d

276 recombinant Aurora B-INCENP inhibits Xenopus MCAK activity in vitro in a phosphorylation-dependent ma

277 the microtubule-depolymerizing kinesin XKCM1/MCAK.