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J Mol Biol. 2019 Aug 17. pii: S0022-2836(19)30506-6. doi: 10.1016/j.jmb.2019.08.003. [Epub ahead of print]

Structure of the Centromere Binding Factor 3 Complex from Kluyveromyces lactis.

Author information

1
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA; Graduate Program in Virology, Harvard Medical School, Boston, MA, 02115, USA.
2
The National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, 10027, USA.
3
Department of Pharmacological Sciences, Stony Brook University School of Medicine, Stony Brook, NY, 11794, USA.
4
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, 02115, USA; Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, 02115, USA. Electronic address: harrison@crystal.harvard.edu.

Abstract

Kinetochores are the multiprotein complexes that link chromosomal centromeres to mitotic-spindle microtubules. Budding yeast centromeres comprise three sequential "centromere-determining elements", CDEI, II, and III. CDEI (8 bp) and CDEIII (~25 bp) are conserved between Kluyveromyces lactis and Saccharomyces cerevisiae, but CDEII in the former is twice as long (160 bp) as CDEII in the latter (80 bp). The CBF3 complex recognizes CDEIII and is required for assembly of a centromeric nucleosome, which in turn recruits other kinetochore components. To understand differences in centromeric nucleosome assembly between K. lactis and S. cerevisiae, we determined the structure of a K. lactis CBF3 complex by electron cryomicroscopy at ~4 Å resolution and compared it with published structures of S. cerevisiae CBF3. We show differences in the pose of Ndc10 and discuss potential models of the K. lactis centromeric nucleosome that account for the extended CDEII length.

KEYWORDS:

budding yeast; centromeric nucleosome assembly; electron cryomicroscopy; kinetochore; point centromere

PMID:
31425683
DOI:
10.1016/j.jmb.2019.08.003

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