Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore
Kinetochore (KTs) are macromolecular protein assemblies that attach sister chromatids to spindle microtubules (MTs) and mediate accurate chromosome segregation during mitosis. The outer KT consists of the KMN network, a protein super-complex comprising Knl1 (yeast Spc105), Mis12 (yeast Mtw1), and Nd...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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The Royal Society
2022-01-01
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| Series: | Open Biology |
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| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsob.210274 |
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| author | Babhrubahan Roy Janice Sim Simon J. Y. Han Ajit P. Joglekar |
| author_facet | Babhrubahan Roy Janice Sim Simon J. Y. Han Ajit P. Joglekar |
| author_sort | Babhrubahan Roy |
| collection | DOAJ |
| description | Kinetochore (KTs) are macromolecular protein assemblies that attach sister chromatids to spindle microtubules (MTs) and mediate accurate chromosome segregation during mitosis. The outer KT consists of the KMN network, a protein super-complex comprising Knl1 (yeast Spc105), Mis12 (yeast Mtw1), and Ndc80 (yeast Ndc80), which harbours sites for MT binding. Within the KMN network, Spc105 acts as an interaction hub of components involved in spindle assembly checkpoint (SAC) signalling. It is known that Spc105 forms a complex with KT component Kre28. However, where Kre28 physically localizes in the budding yeast KT is not clear. The exact function of Kre28 at the KT is also unknown. Here, we investigate how Spc105 and Kre28 interact and how they are organized within bioriented yeast KTs using genetics and cell biological experiments. Our microscopy data show that Spc105 and Kre28 localize at the KT with a 1 : 1 stoichiometry. We also show that the Kre28–Spc105 interaction is important for Spc105 protein turn-over and essential for their mutual recruitment at the KTs. We created several truncation mutants of kre28 that affect Spc105 loading at the KTs. When over-expressed, these mutants sustain the cell viability, but SAC signalling and KT biorientation are impaired. Therefore, we conclude that Kre28 contributes to chromosome biorientation and high-fidelity segregation at least indirectly by regulating Spc105 localization at the KTs. |
| first_indexed | 2024-03-13T07:51:00Z |
| format | Article |
| id | doaj.art-56c1a387fcf14bb9b601a90474089e51 |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-03-13T07:51:00Z |
| publishDate | 2022-01-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-56c1a387fcf14bb9b601a90474089e512023-06-02T16:06:13ZengThe Royal SocietyOpen Biology2046-24412022-01-0112110.1098/rsob.210274Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochoreBabhrubahan Roy0Janice Sim1Simon J. Y. Han2Ajit P. Joglekar3Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USACell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USACell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USACell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USAKinetochore (KTs) are macromolecular protein assemblies that attach sister chromatids to spindle microtubules (MTs) and mediate accurate chromosome segregation during mitosis. The outer KT consists of the KMN network, a protein super-complex comprising Knl1 (yeast Spc105), Mis12 (yeast Mtw1), and Ndc80 (yeast Ndc80), which harbours sites for MT binding. Within the KMN network, Spc105 acts as an interaction hub of components involved in spindle assembly checkpoint (SAC) signalling. It is known that Spc105 forms a complex with KT component Kre28. However, where Kre28 physically localizes in the budding yeast KT is not clear. The exact function of Kre28 at the KT is also unknown. Here, we investigate how Spc105 and Kre28 interact and how they are organized within bioriented yeast KTs using genetics and cell biological experiments. Our microscopy data show that Spc105 and Kre28 localize at the KT with a 1 : 1 stoichiometry. We also show that the Kre28–Spc105 interaction is important for Spc105 protein turn-over and essential for their mutual recruitment at the KTs. We created several truncation mutants of kre28 that affect Spc105 loading at the KTs. When over-expressed, these mutants sustain the cell viability, but SAC signalling and KT biorientation are impaired. Therefore, we conclude that Kre28 contributes to chromosome biorientation and high-fidelity segregation at least indirectly by regulating Spc105 localization at the KTs.https://royalsocietypublishing.org/doi/10.1098/rsob.210274kinetochoremicrotubulespindle assembly checkpoint |
| spellingShingle | Babhrubahan Roy Janice Sim Simon J. Y. Han Ajit P. Joglekar Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore Open Biology kinetochore microtubule spindle assembly checkpoint |
| title | Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore |
| title_full | Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore |
| title_fullStr | Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore |
| title_full_unstemmed | Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore |
| title_short | Kre28–Spc105 interaction is essential for Spc105 loading at the kinetochore |
| title_sort | kre28 spc105 interaction is essential for spc105 loading at the kinetochore |
| topic | kinetochore microtubule spindle assembly checkpoint |
| url | https://royalsocietypublishing.org/doi/10.1098/rsob.210274 |
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