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PMID: 16785321 Published · ppublish English Journal Article

Structural role of Sfi1p-centrin filaments in budding yeast spindle pole body duplication.

The Journal of cell biology ·Vol. 173 ·No. 6 ·2006-06-19 ·Pages 867-77

Li S, Sandercock AM, Conduit P, Robinson CV, Williams RL, Kilmartin JV

Abstract

Centrins are calmodulin-like proteins present in centrosomes and yeast spindle pole bodies (SPBs) and have essential functions in their duplication. The Saccharomyces cerevisiae centrin, Cdc31p, binds Sfi1p on multiple conserved repeats; both proteins localize to the SPB half-bridge, where the new SPB is assembled. The crystal structures of Sfi1p-centrin complexes containing several repeats show Sfi1p as an alpha helix with centrins wrapped around each repeat and similar centrin-centrin contacts between each repeat. Electron microscopy (EM) shadowing of an Sfi1p-centrin complex with 15 Sfi1 repeats and 15 centrins bound showed filaments 60 nm long, compatible with all the Sfi1 repeats as a continuous alpha helix. Immuno-EM localization of the Sfi1p N and C termini showed Sfi1p-centrin filaments spanning the length of the half-bridge with the Sfi1p N terminus at the SPB. This suggests a model for SPB duplication where the half-bridge doubles in length by association of the Sfi1p C termini, thereby providing a new Sfi1p N terminus to initiate SPB assembly.

MeSH Terms
Binding Sites Calcium-Binding Proteins/chemistry,metabolism,physiology Cell Cycle Proteins/chemistry,metabolism,physiology Crystallography, X-Ray Mass Spectrometry Microtubule Proteins/chemistry,metabolism,physiology Models, Molecular Protein Binding Protein Structure, Tertiary Repetitive Sequences, Amino Acid Repressor Proteins/chemistry,metabolism,physiology Saccharomyces cerevisiae/metabolism,ultrastructure Saccharomyces cerevisiae Proteins/chemistry,metabolism,physiology Sequence Analysis, Protein Spindle Apparatus/physiology,ultrastructure
Chemicals
CDC31 protein, S cerevisiae Calcium-Binding Proteins Cell Cycle Proteins Microtubule Proteins Repressor Proteins SFI1 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Li Sam
Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, England, UK.
Sandercock Alan M
Conduit Paul
Robinson Carol V
Williams Roger L
Kilmartin John V
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200603153
Published
2006-06-19
Pages
867-77
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2063913
Subset
IM
Grants
Medical Research Council · MC_U105184282 · United Kingdom
Medical Research Council · MC_U105184308 · United Kingdom
External Links
PubMed source DOI: 10.1083/jcb.200603153 PMC Full Text View journal details
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