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Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface

The conserved Ndc80 kinetochore complex, Ndc80c, is the principal link between mitotic spindle microtubules and centromere-associated proteins. We used AlphaFold 2 (AF2) to obtain predictions of the Ndc80 ‘loop’ structure and of the Ndc80 : Nuf2 globular head domains that interact with the Dam1 subu...

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Autores principales: Zahm, Jacob A., Jenni, Simon, Harrison, Stephen C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993044/
https://www.ncbi.nlm.nih.gov/pubmed/36883282
http://dx.doi.org/10.1098/rsob.220378
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author Zahm, Jacob A.
Jenni, Simon
Harrison, Stephen C.
author_facet Zahm, Jacob A.
Jenni, Simon
Harrison, Stephen C.
author_sort Zahm, Jacob A.
collection PubMed
description The conserved Ndc80 kinetochore complex, Ndc80c, is the principal link between mitotic spindle microtubules and centromere-associated proteins. We used AlphaFold 2 (AF2) to obtain predictions of the Ndc80 ‘loop’ structure and of the Ndc80 : Nuf2 globular head domains that interact with the Dam1 subunit of the heterodecameric DASH/Dam1 complex (Dam1c). The predictions guided design of crystallizable constructs, with structures close to the predicted ones. The Ndc80 ‘loop’ is a stiff, α-helical ‘switchback’ structure; AF2 predictions and positions of preferential cleavage sites indicate that flexibility within the long Ndc80c rod occurs instead at a hinge closer to the globular head. Conserved stretches of the Dam1 C terminus bind Ndc80c such that phosphorylation of Dam1 serine residues 257, 265 and 292 by the mitotic kinase Ipl1/Aurora B can release this contact during error correction of mis-attached kinetochores. We integrate the structural results presented here into our current molecular model of the kinetochore–microtubule interface. The model illustrates how multiple interactions between Ndc80c, DASH/Dam1c and the microtubule lattice stabilize kinetochore attachments.
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spelling pubmed-99930442023-03-09 Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface Zahm, Jacob A. Jenni, Simon Harrison, Stephen C. Open Biol Research The conserved Ndc80 kinetochore complex, Ndc80c, is the principal link between mitotic spindle microtubules and centromere-associated proteins. We used AlphaFold 2 (AF2) to obtain predictions of the Ndc80 ‘loop’ structure and of the Ndc80 : Nuf2 globular head domains that interact with the Dam1 subunit of the heterodecameric DASH/Dam1 complex (Dam1c). The predictions guided design of crystallizable constructs, with structures close to the predicted ones. The Ndc80 ‘loop’ is a stiff, α-helical ‘switchback’ structure; AF2 predictions and positions of preferential cleavage sites indicate that flexibility within the long Ndc80c rod occurs instead at a hinge closer to the globular head. Conserved stretches of the Dam1 C terminus bind Ndc80c such that phosphorylation of Dam1 serine residues 257, 265 and 292 by the mitotic kinase Ipl1/Aurora B can release this contact during error correction of mis-attached kinetochores. We integrate the structural results presented here into our current molecular model of the kinetochore–microtubule interface. The model illustrates how multiple interactions between Ndc80c, DASH/Dam1c and the microtubule lattice stabilize kinetochore attachments. The Royal Society 2023-03-08 /pmc/articles/PMC9993044/ /pubmed/36883282 http://dx.doi.org/10.1098/rsob.220378 Text en © 2023 The Authors. https://creativecommons.org/licenses/by/4.0/Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, provided the original author and source are credited.
spellingShingle Research
Zahm, Jacob A.
Jenni, Simon
Harrison, Stephen C.
Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title_full Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title_fullStr Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title_full_unstemmed Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title_short Structure of the Ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
title_sort structure of the ndc80 complex and its interactions at the yeast kinetochore–microtubule interface
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993044/
https://www.ncbi.nlm.nih.gov/pubmed/36883282
http://dx.doi.org/10.1098/rsob.220378
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