Structure of the DASH/Dam1 complex shows its role at the yeast kinetochore-microtubule interface

Jenni, Simon; Harrison, Stephen C.

doi:10.1126/science.aar6436

Structure of the DASH/Dam1 complex shows its role at the yeast kinetochore-microtubule interface

Journal Article · Fri May 04 00:00:00 EDT 2018 · Science

DOI:https://doi.org/10.1126/science.aar6436· OSTI ID:1438840

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Harvard Medical School, Boston, MA (United States)
Harvard Medical School, Boston, MA (United States); Howard Hughes Medical Inst., Chevy Chase, MD (United States)

Kinetochores connect mitotic-spindle microtubules with chromosomes, allowing microtubule depolymerization to pull chromosomes apart during anaphase while resisting detachment as the microtubule shortens. The heterodecameric DASH/Dam1 complex (DASH/Dam1c), an essential component of yeast kinetochores, assembles into a microtubule-encircling ring. The ring associates with rodlike Ndc80 complexes to organize the kinetochore-microtubule interface. Here, we report the cryo–electron microscopy structure (at ~4.5-angstrom resolution) of a DASH/Dam1c ring and a molecular model of its ordered components, validated by evolutionary direct-coupling analysis. Integrating this structure with that of the Ndc80 complex and with published interaction data yields a molecular picture of kinetochore-microtubule attachment, including how flexible, C-terminal extensions of DASH/Dam1c subunits project and contact widely separated sites on the Ndc80 complex rod and how phosphorylation at previously identified sites might regulate kinetochore assembly.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: Damon Runyon Cancer Research Foundation; National Institute of Health (NIH); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1438840

Journal Information:: Science, Journal Name: Science Journal Issue: 6388 Vol. 360; ISSN 0036-8075

Publisher:: AAASCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Improved contact prediction in proteins: Using pseudolikelihoods to infer Potts models Ekeberg, Magnus; Lövkvist, Cecilia; Lan, Yueheng arXiv https://doi.org/10.48550/arxiv.1211.1281	text	January 2012
Fast pseudolikelihood maximization for direct-coupling analysis of protein structure from many homologous amino-acid sequences Ekeberg, Magnus; Hartonen, Tuomo; Aurell, Erik arXiv https://doi.org/10.48550/arxiv.1401.4832	text	January 2014
Measuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6 Grant, Timothy; Grigorieff, Nikolaus eLife, Vol. 4 https://doi.org/10.7554/eLife.06980	journal	May 2015
Molecular basis of outer kinetochore assembly on CENP-T Huis in 't Veld, Pim J.; Jeganathan, Sadasivam; Petrovic, Arsen eLife, Vol. 5 https://doi.org/10.7554/eLife.21007	journal	December 2016
The Ndc80 complex bridges two Dam1 complex rings Kim, Jae ook; Zelter, Alex; Umbreit, Neil T. eLife, Vol. 6 https://doi.org/10.7554/eLife.21069	journal	February 2017

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The stoichiometry of the outer kinetochore is modulated by microtubule-proximal regulatory factors Dhatchinamoorthy, Karthik; Unruh, Jay R.; Lange, Jeffrey J. Journal of Cell Biology, Vol. 218, Issue 7 https://doi.org/10.1083/jcb.201810070	journal	May 2019
Extensive loss of cell-cycle and DNA repair genes in an ancient lineage of bipolar budding yeasts Steenwyk, Jacob L.; Opulente, Dana A.; Kominek, Jacek PLOS Biology, Vol. 17, Issue 5 https://doi.org/10.1371/journal.pbio.3000255	journal	May 2019
Outer kinetochore protein Dam1 promotes centromere clustering in parallel with Slk19 in budding yeast Mittal, Priyanka; Chavan, Ankita; Trakroo, Deepika Chromosoma, Vol. 128, Issue 2 https://doi.org/10.1007/s00412-019-00694-9	journal	March 2019
Glucose Signaling Is Connected to Chromosome Segregation Through Protein Kinase A Phosphorylation of the Dam1 Kinetochore Subunit in Saccharomyces cerevisiae Shah, Sameer B.; Parmiter, David; Constantine, Christian Genetics, Vol. 211, Issue 2 https://doi.org/10.1534/genetics.118.301727	journal	December 2018

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