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PDBsum entry 2ve7
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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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References listed in PDB file
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Key reference
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Title
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Implications for kinetochore-Microtubule attachment from the structure of an engineered ndc80 complex.
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Authors
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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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Ref.
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Cell, 2008,
133,
427-439.
[DOI no: ]
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PubMed id
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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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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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