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Contents |
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246 a.a.
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303 a.a.
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219 a.a.
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242 a.a.
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* Residue conservation analysis
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PDB id:
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Cell cycle
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Title:
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Crystal structure of a bonsai version of the human ndc80 complex
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Structure:
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Kinetochore protein hec1, kinetochore protein spc25. Chain: a, b. Fragment: chimera of ndc80 residues 80-286 with spc25 residues 118- 224. Synonym: ndc80-spc25 chimera, hshec1, kinetochore-associated protein 2, highly expressed in cancer protein, retinoblastoma-associated protein hec, hspc25. Engineered: yes. Other_details: peptide link between resi 286 and resi 1118.
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Source:
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Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 469008.
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Resolution:
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2.88Å
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R-factor:
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0.234
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R-free:
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0.261
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Authors:
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C.Ciferri,S.Pasqualato,G.Dos Reis,E.Screpanti,A.Maiolica,J.Polka, J.G.De Luca,P.De Wulf,M.Salek,J.Rappsilber,C.A.Moores,E.D.Salmon, A.Musacchio
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Key ref:
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C.Ciferri
et al.
(2008).
Implications for kinetochore-microtubule attachment from the structure of an engineered Ndc80 complex.
Cell,
133,
427-439.
PubMed id:
DOI:
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Date:
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17-Oct-07
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Release date:
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13-May-08
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PROCHECK
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Headers
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References
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O14777
(NDC80_HUMAN) -
Kinetochore protein NDC80 homolog from Homo sapiens
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Seq:
Struc:
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642 a.a.
246 a.a.*
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Q9HBM1
(SPC25_HUMAN) -
Kinetochore protein Spc25 from Homo sapiens
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Seq:
Struc:
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224 a.a.
246 a.a.*
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O14777
(NDC80_HUMAN) -
Kinetochore protein NDC80 homolog from Homo sapiens
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Seq:
Struc:
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642 a.a.
303 a.a.*
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Q9HBM1
(SPC25_HUMAN) -
Kinetochore protein Spc25 from Homo sapiens
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Seq:
Struc:
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224 a.a.
303 a.a.*
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Q8NBT2
(SPC24_HUMAN) -
Kinetochore protein Spc24 from Homo sapiens
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Seq:
Struc:
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197 a.a.
219 a.a.*
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Q9BZD4
(NUF2_HUMAN) -
Kinetochore protein Nuf2 from Homo sapiens
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Seq:
Struc:
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464 a.a.
219 a.a.*
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Enzyme class:
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Chains A, B, C, D:
E.C.?
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DOI no:
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Cell
133:427-439
(2008)
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PubMed id:
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Implications for kinetochore-microtubule attachment from the structure of an engineered Ndc80 complex.
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C.Ciferri,
S.Pasqualato,
E.Screpanti,
G.Varetti,
S.Santaguida,
G.Dos Reis,
A.Maiolica,
J.Polka,
J.G.De Luca,
P.De Wulf,
M.Salek,
J.Rappsilber,
C.A.Moores,
E.D.Salmon,
A.Musacchio.
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ABSTRACT
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Kinetochores are proteinaceous assemblies that mediate the interaction of
chromosomes with the mitotic spindle. The 180 kDa Ndc80 complex is a direct
point of contact between kinetochores and microtubules. Its four subunits
contain coiled coils and form an elongated rod structure with functional
globular domains at either end. We crystallized an engineered "bonsai" Ndc80
complex containing a shortened rod domain but retaining the globular domains
required for kinetochore localization and microtubule binding. The structure
reveals a microtubule-binding interface containing a pair of tightly interacting
calponin-homology (CH) domains with a previously unknown arrangement. The
interaction with microtubules is cooperative and predominantly electrostatic. It
involves positive charges in the CH domains and in the N-terminal tail of the
Ndc80 subunit and negative charges in tubulin C-terminal tails and is regulated
by the Aurora B kinase. We discuss our results with reference to current models
of kinetochore-microtubule attachment and centromere organization.
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Selected figure(s)
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Figure 3.
Figure 3. Organization of the CH Domains in the Ndc80-Nuf2
Subcomplex
(A) Topology diagram of Ndc80 and Nuf2. The CH
domain is contained between the αA and αG helices.
(B)
Two views of the superposition of the CH domains of Ndc80 and
Nuf2. Note the conspicuous bending of the tips of the Nuf2 αE
helix.
(C) Structure-based sequence alignment of the CH
domains of Ndc80, Nuf2, and EB1. The αA, αC, αE, and αG
helices are contoured. Residues highlighted in gray have their
side chains buried in the hydrophobic core of the CH domain.
(D) General view and closeups of the interface of Ndc80 and
Nuf2.
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Figure 6.
Figure 6. Models of Ndc80 and Microtubule-Kinetochore
Interaction
(A–D) Models of the Ndc80-microtubule
interaction. A yellow patch on tubulin in (A) and (B) represents
a hypothetical binding site for the Ndc80 N-terminal tail. In
(C), it is hypothesized that the N-terminal tail binds to the
negatively charged patch on Nuf2, shown in Figure 4B.
(E)
The Ndc80^bonsai complex.
(F) Summary of crosslinking
analysis (Maiolica et al., 2007). Connected black dots mark
crosslinked residues. Numbers in hexagons define distances
between “milestones,” such as subsequent crosslinked
residues or pairs of interacting residues identified in the
structure.
(G) Model of the full-length Ndc80 complex
showing the predicted break in the coiled-coil region.
(H)
Implications from the structure of the Ndc80 complex on the
organization of the microtubule-kinetochore interface.
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The above figures are
reprinted
by permission from Cell Press:
Cell
(2008,
133,
427-439)
copyright 2008.
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Figures were
selected
by an automated process.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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A.Schleiffer,
M.Maier,
G.Litos,
F.Lampert,
P.Hornung,
K.Mechtler,
and
S.Westermann
(2012).
CENP-T proteins are conserved centromere receptors of the Ndc80 complex.
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| |
Nat Cell Biol,
14,
604-613.
|
|
|
|
|
|
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D.Varma,
S.Chandrasekaran,
L.J.Sundin,
K.T.Reidy,
X.Wan,
D.A.Chasse,
K.R.Nevis,
J.G.DeLuca,
E.D.Salmon,
and
J.G.Cook
(2012).
Recruitment of the human Cdt1 replication licensing protein by the loop domain of Hec1 is required for stable kinetochore-microtubule attachment.
|
| |
Nat Cell Biol,
14,
593-603.
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|
|
|
|
|
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E.A.Foley,
and
T.M.Kapoor
(2012).
Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore.
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| |
Nat Rev Mol Cell Biol,
14,
25-37.
|
|
|
|
|
|
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G.M.Alushin,
V.Musinipally,
D.Matson,
J.Tooley,
P.T.Stukenberg,
and
E.Nogales
(2012).
Multimodal microtubule binding by the Ndc80 kinetochore complex.
|
| |
Nat Struct Mol Biol,
19,
1161-1167.
|
|
|
|
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|
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A.M.Olszak,
D.van Essen,
A.J.Pereira,
S.Diehl,
T.Manke,
H.Maiato,
S.Saccani,
and
P.Heun
(2011).
Heterochromatin boundaries are hotspots for de novo kinetochore formation.
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| |
Nat Cell Biol,
13,
799-808.
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|
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C.L.Asbury,
J.F.Tien,
and
T.N.Davis
(2011).
Kinetochores' gripping feat: conformational wave or biased diffusion?
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| |
Trends Cell Biol,
21,
38-46.
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|
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|
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|
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F.Lampert,
and
S.Westermann
(2011).
A blueprint for kinetochores - new insights into the molecular mechanics of cell division.
|
| |
Nat Rev Mol Cell Biol,
12,
407-412.
|
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|
|
|
|
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H.Zhang,
and
R.K.Dawe
(2011).
Mechanisms of plant spindle formation.
|
| |
Chromosome Res,
19,
335-344.
|
|
|
|
|
|
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J.C.Schmidt,
and
I.M.Cheeseman
(2011).
Chromosome segregation: keeping kinetochores in the loop.
|
| |
Curr Biol,
21,
R110-R112.
|
|
|
|
|
|
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J.F.Maure,
S.Komoto,
Y.Oku,
A.Mino,
S.Pasqualato,
K.Natsume,
L.Clayton,
A.Musacchio,
and
T.U.Tanaka
(2011).
The Ndc80 loop region facilitates formation of kinetochore attachment to the dynamic microtubule plus end.
|
| |
Curr Biol,
21,
207-213.
|
|
|
|
|
|
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J.Tooley,
and
P.T.Stukenberg
(2011).
The Ndc80 complex: integrating the kinetochore's many movements.
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| |
Chromosome Res,
19,
377-391.
|
|
|
|
|
|
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K.E.Gascoigne,
K.Takeuchi,
A.Suzuki,
T.Hori,
T.Fukagawa,
and
I.M.Cheeseman
(2011).
Induced Ectopic Kinetochore Assembly Bypasses the Requirement for CENP-A Nucleosomes.
|
| |
Cell,
145,
410-422.
|
|
|
|
|
|
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K.S.Hsu,
and
T.Toda
(2011).
Ndc80 internal loop interacts with Dis1/TOG to ensure proper kinetochore-spindle attachment in fission yeast.
|
| |
Curr Biol,
21,
214-220.
|
|
|
|
|
|
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M.A.Lampson,
and
I.M.Cheeseman
(2011).
Sensing centromere tension: Aurora B and the regulation of kinetochore function.
|
| |
Trends Cell Biol,
21,
133-140.
|
|
|
|
|
|
|
P.O.Widlund,
J.H.Stear,
A.Pozniakovsky,
M.Zanic,
S.Reber,
G.J.Brouhard,
A.A.Hyman,
and
J.Howard
(2011).
XMAP215 polymerase activity is built by combining multiple tubulin-binding TOG domains and a basic lattice-binding region.
|
| |
Proc Natl Acad Sci U S A,
108,
2741-2746.
|
|
|
|
|
|
|
A.K.Caydasi,
B.Ibrahim,
and
G.Pereira
(2010).
Monitoring spindle orientation: Spindle position checkpoint in charge.
|
| |
Cell Div,
5,
28.
|
|
|
|
|
|
|
A.Leitner,
T.Walzthoeni,
A.Kahraman,
F.Herzog,
O.Rinner,
M.Beck,
and
R.Aebersold
(2010).
Probing native protein structures by chemical cross-linking, mass spectrometry, and bioinformatics.
|
| |
Mol Cell Proteomics,
9,
1634-1649.
|
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|
|
|
|
|
A.P.Carter,
and
R.D.Vale
(2010).
Communication between the AAA+ ring and microtubule-binding domain of dynein.
|
| |
Biochem Cell Biol,
88,
15-21.
|
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|
|
A.Petrovic,
S.Pasqualato,
P.Dube,
V.Krenn,
S.Santaguida,
D.Cittaro,
S.Monzani,
L.Massimiliano,
J.Keller,
A.Tarricone,
A.Maiolica,
H.Stark,
and
A.Musacchio
(2010).
The MIS12 complex is a protein interaction hub for outer kinetochore assembly.
|
| |
J Cell Biol,
190,
835-852.
|
|
|
|
|
|
|
A.R.Cole,
L.P.Lewis,
and
H.Walden
(2010).
The structure of the catalytic subunit FANCL of the Fanconi anemia core complex.
|
| |
Nat Struct Mol Biol,
17,
294-298.
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PDB code:
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B.F.McEwen,
and
Y.Dong
(2010).
Contrasting models for kinetochore microtubule attachment in mammalian cells.
|
| |
Cell Mol Life Sci,
67,
2163-2172.
|
|
|
|
|
|
|
C.Rumpf,
L.Cipak,
A.Schleiffer,
A.Pidoux,
K.Mechtler,
I.M.Tolić-Nørrelykke,
and
J.Gregan
(2010).
Laser microsurgery provides evidence for merotelic kinetochore attachments in fission yeast cells lacking Pcs1 or Clr4.
|
| |
Cell Cycle,
9,
3997-4004.
|
|
|
|
|
|
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D.Liu,
M.Vleugel,
C.B.Backer,
T.Hori,
T.Fukagawa,
I.M.Cheeseman,
and
M.A.Lampson
(2010).
Regulated targeting of protein phosphatase 1 to the outer kinetochore by KNL1 opposes Aurora B kinase.
|
| |
J Cell Biol,
188,
809-820.
|
|
|
|
|
|
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D.P.Maskell,
X.W.Hu,
and
M.R.Singleton
(2010).
Molecular architecture and assembly of the yeast kinetochore MIND complex.
|
| |
J Cell Biol,
190,
823-834.
|
|
|
|
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|
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E.Screpanti,
S.Santaguida,
T.Nguyen,
R.Silvestri,
R.Gussio,
A.Musacchio,
E.Hamel,
and
P.De Wulf
(2010).
A screen for kinetochore-microtubule interaction inhibitors identifies novel antitubulin compounds.
|
| |
PLoS One,
5,
e11603.
|
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|
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F.Lampert,
P.Hornung,
and
S.Westermann
(2010).
The Dam1 complex confers microtubule plus end-tracking activity to the Ndc80 kinetochore complex.
|
| |
J Cell Biol,
189,
641-649.
|
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|
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G.Civelekoglu-Scholey,
and
J.M.Scholey
(2010).
Mitotic force generators and chromosome segregation.
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| |
Cell Mol Life Sci,
67,
2231-2250.
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G.J.Kops,
A.T.Saurin,
and
P.Meraldi
(2010).
Finding the middle ground: how kinetochores power chromosome congression.
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| |
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67,
2145-2161.
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|
|
G.M.Alushin,
V.H.Ramey,
S.Pasqualato,
D.A.Ball,
N.Grigorieff,
A.Musacchio,
and
E.Nogales
(2010).
The Ndc80 kinetochore complex forms oligomeric arrays along microtubules.
|
| |
Nature,
467,
805-810.
|
|
|
PDB code:
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|
|
H.Foussard,
P.Ferrer,
P.Valenti,
C.Polesello,
S.Carreno,
and
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| |
PLoS One,
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H.Maiato,
and
M.Lince-Faria
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(2010).
The monopolin complex crosslinks kinetochore components to regulate chromosome-microtubule attachments.
|
| |
Cell,
142,
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PDB codes:
|
|
|
|
|
|
|
|
L.J.Sundin,
and
J.G.Deluca
(2010).
Kinetochores: NDC80 toes the line.
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| |
Curr Biol,
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Dissecting the role of MPS1 in chromosome biorientation and the spindle checkpoint through the small molecule inhibitor reversine.
|
| |
J Cell Biol,
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(2010).
Kinetochore-microtubule interactions: steps towards bi-orientation.
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