C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast

Medini Ghodgaonkar-Steger; Mia Potocnjak; Tomasz Zimniak; Josef Fischböck-Halwachs; Victor Solis-Mezarino; Sylvia Singh; Tea Speljko; Götz Hagemann; David Jan Drexler; Gregor Witte; Franz Herzog

doi:10.1016/j.celrep.2020.108190

C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast

Medini Ghodgaonkar-Steger, Mia Potocnjak, Tomasz Zimniak, Josef Fischböck-Halwachs, Victor Solis-Mezarino, Sylvia Singh, Tea Speljko, Götz Hagemann, David Jan Drexler, Gregor Witte, Franz Herzog

Research output: Contribution to journal › Article › peer-review

Abstract

Kinetochores are macromolecular protein assemblies at centromeres that mediate accurate chromosome segregation during cell division. The outer kinetochore KNL1 SPC105, MIS12 MTW1, and NDC80 NDC80 complexes assemble the KMN network, which harbors the sites of microtubule binding and spindle assembly checkpoint signaling. The buildup of the KMN network that transmits microtubule pulling forces to budding yeast point centromeres is poorly understood. Here, we identify 225 inter-protein crosslinks by mass spectrometry on KMN complexes isolated from Saccharomyces cerevisiae that delineate the KMN subunit connectivity for outer kinetochore assembly. C-Terminal motifs of Nsl1 and Mtw1 recruit the SPC105 complex through Kre28, and both motifs aid tethering of the NDC80 complex by the previously reported Dsn1 C terminus. We show that a hub of three C-terminal MTW1 subunit motifs mediates the cooperative stabilization of the KMN network, which is augmented by a direct NDC80-SPC105 association.

Original language	English
Article number	108190
Pages (from-to)	108190
Journal	Cell Reports
Volume	32
Issue number	13
DOIs	https://doi.org/10.1016/j.celrep.2020.108190
Publication status	Published - 29 Sept 2020
Externally published	Yes

Keywords

connectivity
cooperativity
crosslinking
kinetochore
KMN network
mass spectrometry
microtubule binding
MTW1
NDC80
SPC105
Kinetochores/metabolism
Amino Acid Sequence
Saccharomyces cerevisiae/metabolism
Mass Spectrometry/methods
Saccharomycetales/pathogenicity
Microtubule-Associated Proteins/metabolism

Research fields

Cell Division
Chemical Crosslinking
Mass spectrometry
Structural Proteomics

IMC Research Focuses

Medical biotechnology

ÖFOS 2012 - Austrian Fields of Study

106037 Proteomics
106041 Structural biology
106044 Systems biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.celrep.2020.108190

Cite this

Ghodgaonkar-Steger, M., Potocnjak, M., Zimniak, T., Fischböck-Halwachs, J., Solis-Mezarino, V., Singh, S., Speljko, T., Hagemann, G., Drexler, D. J., Witte, G., & Herzog, F. (2020). C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast. Cell Reports, 32(13), 108190. Article 108190. https://doi.org/10.1016/j.celrep.2020.108190

@article{1d9264cbddf640db96378c439f28b943,

title = "C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast",

abstract = "Kinetochores are macromolecular protein assemblies at centromeres that mediate accurate chromosome segregation during cell division. The outer kinetochore KNL1 SPC105, MIS12 MTW1, and NDC80 NDC80 complexes assemble the KMN network, which harbors the sites of microtubule binding and spindle assembly checkpoint signaling. The buildup of the KMN network that transmits microtubule pulling forces to budding yeast point centromeres is poorly understood. Here, we identify 225 inter-protein crosslinks by mass spectrometry on KMN complexes isolated from Saccharomyces cerevisiae that delineate the KMN subunit connectivity for outer kinetochore assembly. C-Terminal motifs of Nsl1 and Mtw1 recruit the SPC105 complex through Kre28, and both motifs aid tethering of the NDC80 complex by the previously reported Dsn1 C terminus. We show that a hub of three C-terminal MTW1 subunit motifs mediates the cooperative stabilization of the KMN network, which is augmented by a direct NDC80-SPC105 association. ",

keywords = "connectivity, cooperativity, crosslinking, kinetochore, KMN network, mass spectrometry, microtubule binding, MTW1, NDC80, SPC105, Kinetochores/metabolism, Amino Acid Sequence, Saccharomyces cerevisiae/metabolism, Mass Spectrometry/methods, Saccharomycetales/pathogenicity, Microtubule-Associated Proteins/metabolism",

author = "Medini Ghodgaonkar-Steger and Mia Potocnjak and Tomasz Zimniak and Josef Fischb{\"o}ck-Halwachs and Victor Solis-Mezarino and Sylvia Singh and Tea Speljko and G{\"o}tz Hagemann and Drexler, {David Jan} and Gregor Witte and Franz Herzog",

year = "2020",

month = sep,

day = "29",

doi = "10.1016/j.celrep.2020.108190",

language = "English",

volume = "32",

pages = "108190",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Cell Press",

number = "13",

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Ghodgaonkar-Steger, M, Potocnjak, M, Zimniak, T, Fischböck-Halwachs, J, Solis-Mezarino, V, Singh, S, Speljko, T, Hagemann, G, Drexler, DJ, Witte, G & Herzog, F 2020, 'C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast', Cell Reports, vol. 32, no. 13, 108190, pp. 108190. https://doi.org/10.1016/j.celrep.2020.108190

TY - JOUR

T1 - C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast

AU - Ghodgaonkar-Steger, Medini

AU - Potocnjak, Mia

AU - Zimniak, Tomasz

AU - Fischböck-Halwachs, Josef

AU - Solis-Mezarino, Victor

AU - Singh, Sylvia

AU - Speljko, Tea

AU - Hagemann, Götz

AU - Drexler, David Jan

AU - Witte, Gregor

AU - Herzog, Franz

PY - 2020/9/29

Y1 - 2020/9/29

N2 - Kinetochores are macromolecular protein assemblies at centromeres that mediate accurate chromosome segregation during cell division. The outer kinetochore KNL1 SPC105, MIS12 MTW1, and NDC80 NDC80 complexes assemble the KMN network, which harbors the sites of microtubule binding and spindle assembly checkpoint signaling. The buildup of the KMN network that transmits microtubule pulling forces to budding yeast point centromeres is poorly understood. Here, we identify 225 inter-protein crosslinks by mass spectrometry on KMN complexes isolated from Saccharomyces cerevisiae that delineate the KMN subunit connectivity for outer kinetochore assembly. C-Terminal motifs of Nsl1 and Mtw1 recruit the SPC105 complex through Kre28, and both motifs aid tethering of the NDC80 complex by the previously reported Dsn1 C terminus. We show that a hub of three C-terminal MTW1 subunit motifs mediates the cooperative stabilization of the KMN network, which is augmented by a direct NDC80-SPC105 association.

AB - Kinetochores are macromolecular protein assemblies at centromeres that mediate accurate chromosome segregation during cell division. The outer kinetochore KNL1 SPC105, MIS12 MTW1, and NDC80 NDC80 complexes assemble the KMN network, which harbors the sites of microtubule binding and spindle assembly checkpoint signaling. The buildup of the KMN network that transmits microtubule pulling forces to budding yeast point centromeres is poorly understood. Here, we identify 225 inter-protein crosslinks by mass spectrometry on KMN complexes isolated from Saccharomyces cerevisiae that delineate the KMN subunit connectivity for outer kinetochore assembly. C-Terminal motifs of Nsl1 and Mtw1 recruit the SPC105 complex through Kre28, and both motifs aid tethering of the NDC80 complex by the previously reported Dsn1 C terminus. We show that a hub of three C-terminal MTW1 subunit motifs mediates the cooperative stabilization of the KMN network, which is augmented by a direct NDC80-SPC105 association.

KW - connectivity

KW - cooperativity

KW - crosslinking

KW - kinetochore

KW - KMN network

KW - mass spectrometry

KW - microtubule binding

KW - MTW1

KW - NDC80

KW - SPC105

KW - Kinetochores/metabolism

KW - Amino Acid Sequence

KW - Saccharomyces cerevisiae/metabolism

KW - Mass Spectrometry/methods

KW - Saccharomycetales/pathogenicity

KW - Microtubule-Associated Proteins/metabolism

UR - http://www.scopus.com/inward/record.url?scp=85091672112&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2020.108190

DO - 10.1016/j.celrep.2020.108190

M3 - Article

C2 - 32997987

AN - SCOPUS:85091672112

SN - 2211-1247

VL - 32

SP - 108190

JO - Cell Reports

JF - Cell Reports

IS - 13

M1 - 108190

ER -

C-Terminal Motifs of the MTW1 Complex Cooperatively Stabilize Outer Kinetochore Assembly in Budding Yeast

Abstract

Keywords

Research fields

IMC Research Focuses

ÖFOS 2012 - Austrian Fields of Study

UN SDGs

Access to Document

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