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Network of protein interactions within the Drosophila inner kinetochore

Network of protein interactions within the Drosophila inner kinetochore
Network of protein interactions within the Drosophila inner kinetochore
The kinetochore provides a physical connection between microtubules and the centromeric regions of chromosomes that is critical for their equitable segregation. The trimeric Mis12 sub-complex of the Drosophila kinetochore binds to the mitotic centromere using CENP-C as a platform. However, knowledge of the precise connections between Mis12 complex components and CENP-C has remained elusive despite the fundamental importance of this part of the cell division machinery. Here, we employ hydrogen-deuterium exchange coupled with mass spectrometry to reveal that Mis12 and Nnf1 form a dimer maintained by interacting coiled-coil (CC) domains within the carboxy-terminal parts of both proteins. Adjacent to these interacting CCs is a carboxy-terminal domain that also interacts with Nsl1. The amino-terminal parts of Mis12 and Nnf1 form a CENP-C-binding surface, which docks the complex and thus the entire kinetochore to mitotic centromeres. Mutational analysis confirms these precise interactions are critical for both structure and function of the complex. Thus, we conclude the organization of the Mis12-Nnf1 dimer confers upon the Mis12 complex a bipolar, elongated structure that is critical for kinetochore function.
1-18
Richter, Magdalena M.
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Poznanski, Jaroslaw
e28e7c5f-db15-439e-ba2f-00f0c115d227
Zdziarska, Anna
063157d3-fb00-45f4-b5cf-577dd892b8a3
Czarnocki-Cieciura, Mariusz
d95e46e8-fc78-4d48-84b5-b84e7382a563
Lipinszki, Zoltan
e405fd6d-1bee-4725-96b7-7c315f2826ed
Dadlez, Michal
c6cc6bee-3459-4449-a2ab-36233c8a0cdc
Glover, David M.
cca9cd19-3e1e-4906-b418-41876c1f9c61
Przewloka, Marcin
9b25e73c-ec15-43df-a5a4-ac9574bb20ab
Richter, Magdalena M.
c65b39a4-87bf-4735-bd08-ec77c5c4ef8b
Poznanski, Jaroslaw
e28e7c5f-db15-439e-ba2f-00f0c115d227
Zdziarska, Anna
063157d3-fb00-45f4-b5cf-577dd892b8a3
Czarnocki-Cieciura, Mariusz
d95e46e8-fc78-4d48-84b5-b84e7382a563
Lipinszki, Zoltan
e405fd6d-1bee-4725-96b7-7c315f2826ed
Dadlez, Michal
c6cc6bee-3459-4449-a2ab-36233c8a0cdc
Glover, David M.
cca9cd19-3e1e-4906-b418-41876c1f9c61
Przewloka, Marcin
9b25e73c-ec15-43df-a5a4-ac9574bb20ab

Richter, Magdalena M., Poznanski, Jaroslaw, Zdziarska, Anna, Czarnocki-Cieciura, Mariusz, Lipinszki, Zoltan, Dadlez, Michal, Glover, David M. and Przewloka, Marcin (2016) Network of protein interactions within the Drosophila inner kinetochore. Open Biology, 6 (2), 1-18. (doi:10.1098/rsob.150238). (PMID:26911623)

Record type: Article

Abstract

The kinetochore provides a physical connection between microtubules and the centromeric regions of chromosomes that is critical for their equitable segregation. The trimeric Mis12 sub-complex of the Drosophila kinetochore binds to the mitotic centromere using CENP-C as a platform. However, knowledge of the precise connections between Mis12 complex components and CENP-C has remained elusive despite the fundamental importance of this part of the cell division machinery. Here, we employ hydrogen-deuterium exchange coupled with mass spectrometry to reveal that Mis12 and Nnf1 form a dimer maintained by interacting coiled-coil (CC) domains within the carboxy-terminal parts of both proteins. Adjacent to these interacting CCs is a carboxy-terminal domain that also interacts with Nsl1. The amino-terminal parts of Mis12 and Nnf1 form a CENP-C-binding surface, which docks the complex and thus the entire kinetochore to mitotic centromeres. Mutational analysis confirms these precise interactions are critical for both structure and function of the complex. Thus, we conclude the organization of the Mis12-Nnf1 dimer confers upon the Mis12 complex a bipolar, elongated structure that is critical for kinetochore function.

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More information

Accepted/In Press date: 1 February 2016
Published date: 24 February 2016
Organisations: Molecular and Cellular, Centre for Biological Sciences

Identifiers

Local EPrints ID: 393853
URI: http://eprints.soton.ac.uk/id/eprint/393853
PURE UUID: 6f8bfb57-2d41-4d2f-b40d-c67ba1e12e70
ORCID for Marcin Przewloka: ORCID iD orcid.org/0000-0002-0329-9162

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Date deposited: 09 May 2016 08:48
Last modified: 17 Dec 2019 01:32

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