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Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein

The kinetochore is the macromolecular machinery that drives chromosome segregation by interacting with spindle microtubules. Kinetoplastids (such as Trypanosoma brucei), a group of evolutionarily divergent eukaryotes, have a unique set of kinetochore proteins that lack any significant homology to ca...

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Detalles Bibliográficos
Autores principales: Ludzia, Patryk, Lowe, Edward D., Marcianò, Gabriele, Mohammed, Shabaz, Redfield, Christina, Akiyoshi, Bungo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8443799/
https://www.ncbi.nlm.nih.gov/pubmed/33915106
http://dx.doi.org/10.1016/j.str.2021.04.004
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author Ludzia, Patryk
Lowe, Edward D.
Marcianò, Gabriele
Mohammed, Shabaz
Redfield, Christina
Akiyoshi, Bungo
author_facet Ludzia, Patryk
Lowe, Edward D.
Marcianò, Gabriele
Mohammed, Shabaz
Redfield, Christina
Akiyoshi, Bungo
author_sort Ludzia, Patryk
collection PubMed
description The kinetochore is the macromolecular machinery that drives chromosome segregation by interacting with spindle microtubules. Kinetoplastids (such as Trypanosoma brucei), a group of evolutionarily divergent eukaryotes, have a unique set of kinetochore proteins that lack any significant homology to canonical kinetochore components. To date, KKT4 is the only kinetoplastid kinetochore protein that is known to bind microtubules. Here we use X-ray crystallography, NMR spectroscopy, and crosslinking mass spectrometry to characterize the structure and dynamics of KKT4. We show that its microtubule-binding domain consists of a coiled-coil structure followed by a positively charged disordered tail. The structure of the C-terminal BRCT domain of KKT4 reveals that it is likely a phosphorylation-dependent protein-protein interaction domain. The BRCT domain interacts with the N-terminal region of the KKT4 microtubule-binding domain and with a phosphopeptide derived from KKT8. Taken together, these results provide structural insights into the unconventional kinetoplastid kinetochore protein KKT4.
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spelling pubmed-84437992021-09-22 Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein Ludzia, Patryk Lowe, Edward D. Marcianò, Gabriele Mohammed, Shabaz Redfield, Christina Akiyoshi, Bungo Structure Article The kinetochore is the macromolecular machinery that drives chromosome segregation by interacting with spindle microtubules. Kinetoplastids (such as Trypanosoma brucei), a group of evolutionarily divergent eukaryotes, have a unique set of kinetochore proteins that lack any significant homology to canonical kinetochore components. To date, KKT4 is the only kinetoplastid kinetochore protein that is known to bind microtubules. Here we use X-ray crystallography, NMR spectroscopy, and crosslinking mass spectrometry to characterize the structure and dynamics of KKT4. We show that its microtubule-binding domain consists of a coiled-coil structure followed by a positively charged disordered tail. The structure of the C-terminal BRCT domain of KKT4 reveals that it is likely a phosphorylation-dependent protein-protein interaction domain. The BRCT domain interacts with the N-terminal region of the KKT4 microtubule-binding domain and with a phosphopeptide derived from KKT8. Taken together, these results provide structural insights into the unconventional kinetoplastid kinetochore protein KKT4. Cell Press 2021-09-02 /pmc/articles/PMC8443799/ /pubmed/33915106 http://dx.doi.org/10.1016/j.str.2021.04.004 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ludzia, Patryk
Lowe, Edward D.
Marcianò, Gabriele
Mohammed, Shabaz
Redfield, Christina
Akiyoshi, Bungo
Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title_full Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title_fullStr Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title_full_unstemmed Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title_short Structural characterization of KKT4, an unconventional microtubule-binding kinetochore protein
title_sort structural characterization of kkt4, an unconventional microtubule-binding kinetochore protein
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8443799/
https://www.ncbi.nlm.nih.gov/pubmed/33915106
http://dx.doi.org/10.1016/j.str.2021.04.004
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