Cargando…
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...
Autores principales: | , , , , , |
---|---|
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 |
_version_ | 1784568365410418688 |
---|---|
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. |
format | Online Article Text |
id | pubmed-8443799 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT ludziapatryk structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein AT loweedwardd structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein AT marcianogabriele structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein AT mohammedshabaz structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein AT redfieldchristina structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein AT akiyoshibungo structuralcharacterizationofkkt4anunconventionalmicrotubulebindingkinetochoreprotein |