ArticleGenetics2013
Coupling unbiased mutagenesis to high-throughput DNA sequencing uncovers functional domains in the Ndc80 kinetochore protein of Saccharomyces cerevisiae.
Article in Genetics, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 21 citations in OpenAlex.
- Structural mechanism of outer kinetochore Dam1-Ndc80 complex assembly on microtubules.Science (New York, N.Y.) · 2023Article
- The Functional Meaning of 5'UTR in Protein-Coding Genes.International journal of molecular sciences · 2023Review
- Three interacting regions of the Ndc80 and Dam1 complexes support microtubule tip-coupling under load.The Journal of cell biology · 2022Article
- Tight bending of the Ndc80 complex provides intrinsic regulation of its binding to microtubules.eLife · 2019Article
- Novel phosphorylation states of the yeast spindle pole body.Biology open · 2018Article
- Measuring mitotic forces.Methods in cell biology · 2018Article
- Tension sensors reveal how the kinetochore shares its load.BioEssays : news and reviews in molecular, cellular and developmental biology · 2017Review
- Article
- Higher-order oligomerization of Spc110p drives γ-tubulin ring complex assembly.Molecular biology of the cell · 2016Article
- The hairpin region of Ndc80 is important for the kinetochore recruitment of Mph1/MPS1 in fission yeast.Cell cycle (Georgetown, Tex.) · 2016Article
- Emerging roles of sumoylation in the regulation of actin, microtubules, intermediate filaments, and septins.Cytoskeleton (Hoboken, N.J.) · 2015Review
- MAPping the Ndc80 loop in cancer: A possible link between Ndc80/Hec1 overproduction and cancer formation.BioEssays : news and reviews in molecular, cellular and developmental biology · 2015Review
- Kinetochore biorientation in Saccharomyces cerevisiae requires a tightly folded conformation of the Ndc80 complex.Genetics · 2014Article
- Force is a signal that cells cannot ignore.Molecular biology of the cell · 2014Review
- Accurate phosphoregulation of kinetochore-microtubule affinity requires unconstrained molecular interactions.The Journal of cell biology · 2014Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
Abstract
During mitosis, kinetochores physically link chromosomes to the dynamic ends of spindle microtubules. This linkage depends on the Ndc80 complex, a conserved and essential microtubule-binding component of the kinetochore. As a member of the complex, the Ndc80 protein forms microtubule attachments through a calponin homology domain. Ndc80 is also required for recruiting other components to the kinetochore and responding to mitotic regulatory signals. While the calponin homology domain has been the focus of biochemical and structural characterization, the function of the remainder of Ndc80 is poorly understood. Here, we utilized a new approach that couples high-throughput sequencing to a saturating linker-scanning mutagenesis screen in Saccharomyces cerevisiae. We identified domains in previously uncharacterized regions of Ndc80 that are essential for its function in vivo. We show that a helical hairpin adjacent to the calponin homology domain influences microtubule binding by the complex. Furthermore, a mutation in this hairpin abolishes the ability of the Dam1 complex to strengthen microtubule attachments made by the Ndc80 complex. Finally, we defined a C-terminal segment of Ndc80 required for tetramerization of the Ndc80 complex in vivo. This unbiased mutagenesis approach can be generally applied to genes in S. cerevisiae to identify functional properties and domains.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.