Structure-function analysis of the C-terminal domain of CNM67, a core component of the Saccharomyces cerevisiae spindle pole body

Vadim A. Klenchin, Jeremiah J. Frye, Michele H. Jones, Mark Winey, Ivan Rayment

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The spindle pole body of the budding yeast Saccharomyces cerevisiae has served as a model system for understanding microtubule organizing centers, yet very little is known about the molecular structure of its components. We report here the structure of the C-terminal domain of the core component Cnm67 at 2.3 Å resolution. The structure determination was aided by a novel approach to crystallization of proteins containing coiled-coils that utilizes globular domains to stabilize the coiled-coils. This enhances their solubility in Escherichia coli and improves their crystallization. The Cnm67 C-terminal domain (residues Asn-429 - Lys-581) exhibits a previously unseen dimeric, interdigitated, all α-helical fold. In vivo studies demonstrate that this domain alone is able to localize to the spindle pole body. In addition, the structure reveals a large functionally indispensable positively charged surface patch that is implicated in spindle pole body localization. Finally, the C-terminal eight residues are disordered but are critical for protein folding and structural stability.

Original languageEnglish (US)
Pages (from-to)18240-18250
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number20
DOIs
StatePublished - May 20 2011
Externally publishedYes

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Spindle Pole Bodies
Yeast
Saccharomyces cerevisiae
Poles
Crystallization
Microtubule-Organizing Center
Protein folding
Saccharomycetales
Protein Folding
Molecular Structure
Solubility
Escherichia coli
Molecular structure
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Structure-function analysis of the C-terminal domain of CNM67, a core component of the Saccharomyces cerevisiae spindle pole body. / Klenchin, Vadim A.; Frye, Jeremiah J.; Jones, Michele H.; Winey, Mark; Rayment, Ivan.

In: Journal of Biological Chemistry, Vol. 286, No. 20, 20.05.2011, p. 18240-18250.

Research output: Contribution to journalArticle

Klenchin, Vadim A. ; Frye, Jeremiah J. ; Jones, Michele H. ; Winey, Mark ; Rayment, Ivan. / Structure-function analysis of the C-terminal domain of CNM67, a core component of the Saccharomyces cerevisiae spindle pole body. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 20. pp. 18240-18250.
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