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Molecular requirements for the formation of a kinetochore–microtubule interface by Dam1 and Ndc80 complexes

Journal Article · · Journal of Cell Biology
 [1];  [2];  [3];  [4];  [2]
  1. Research Inst. of Molecular Pathology, Vienna (Austria); DOE/OSTI
  2. Research Inst. of Molecular Pathology, Vienna (Austria)
  3. Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst. Molecular and Cell Biology Dept. Biophysics Graduate Group
  4. Univ. of California, Berkeley, CA (United States). Howard Hughes Medical Inst. Molecular and Cell Biology Dept. Biophysics Graduate Group; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division
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Kinetochores are large protein complexes that link sister chromatids to the spindle and transduce microtubule dynamics into chromosome movement. In budding yeast, the kinetochore–microtubule interface is formed by the plus end–associated Dam1 complex and the kinetochore-resident Ndc80 complex, but how they work in combination and whether a physical association between them is critical for chromosome segregation is poorly understood. Here, we define structural elements required for the Ndc80–Dam1 interaction and probe their function in vivo. A novel ndc80 allele, selectively impaired in Dam1 binding, displayed growth and chromosome segregation defects. Its combination with an N-terminal truncation resulted in lethality, demonstrating essential but partially redundant roles for the Ndc80 N-tail and Ndc80–Dam1 interface. In contrast, mutations in the calponin homology domain of Ndc80 abrogated kinetochore function and were not compensated by the presence of Dam1. Our experiments shed light on how microtubule couplers cooperate and impose important constraints on structural models for outer kinetochore assembly.
Research Organization:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC)
Sponsoring Organization:
Austrian Science Fund (FWF); European Research Council (ERC); National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1625166
Journal Information:
Journal of Cell Biology, Journal Name: Journal of Cell Biology Journal Issue: 1 Vol. 200; ISSN 0021-9525
Publisher:
Rockefeller University PressCopyright Statement
Country of Publication:
United States
Language:
English

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  • The University of North Carolina at Chapel Hill University Libraries https://doi.org/10.17615/gffh-5h68
text January 2008
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Cited By (22)

Kinetochore–microtubule error correction is driven by differentially regulated interaction modes journal March 2015
How the kinetochore couples microtubule force and centromere stretch to move chromosomes journal March 2016
Kinetochore complexes need teamwork journal December 2012
The internal loop of fission yeast Ndc80 binds Alp7/TACC-Alp14/TOG and ensures proper chromosome attachment journal April 2013
Control of the spindle checkpoint by lateral kinetochore attachment and limited Mad1 recruitment journal July 2015
Stu2 acts as a microtubule destabilizer in metaphase budding yeast spindles journal February 2018
The Agrobacterium VirD5 protein hyperactivates the mitotic Aurora kinase in host cells journal February 2019
Ndc80 Loop as a protein-protein interaction motif journal January 2013
Hec1/Ndc80 Tail Domain Function at the Kinetochore-Microtubule Interface journal February 2020
A Molecular View of Kinetochore Assembly and Function journal January 2017
Non-catalytic motor domains enable processive movement and functional diversification of the kinesin-14 Kar3 journal January 2015
The Ndc80 complex bridges two Dam1 complex rings journal February 2017
Tight bending of the Ndc80 complex provides intrinsic regulation of its binding to microtubules journal May 2019
An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast posted_content August 2018
Coordination of NDC80 and Ska complexes at the kinetochore-microtubule interface in human cells posted_content January 2019
Crosstalk Between Microtubule Attachment Complexes Ensures Accurate Chromosome Segregation journal November 2013
Structure of the DASH/Dam1 complex shows its role at the yeast kinetochore-microtubule interface journal May 2018
The Composition, Functions, and Regulation of the Budding Yeast Kinetochore journal August 2013
Glucose Signaling Is Connected to Chromosome Segregation Through Protein Kinase A Phosphorylation of the Dam1 Kinetochore Subunit in Saccharomyces   cerevisiae journal December 2018
How the kinetochore couples microtubule force and centromere stretch to move chromosomes text January 2016
The kinetochore interaction network (KIN) of ascomycetes journal May 2016
An assay for de novo kinetochore assembly reveals a key role for the CENP-T pathway in budding yeast journal August 2018

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59 BASIC BIOLOGICAL SCIENCES
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