Sfi1p Has Conserved Centrin-Binding Sites and an Essential Function in Budding Yeast Spindle Pole Body Duplication

Centrins are calmodulin-like proteins present in microtubule-organizing centers. The Saccharomyces cerevisiae centrin, Cdc31p, was functionally tagged with a single Z domain of protein A, and used in pull-down experiments to isolate Cdc31p-binding proteins. One of these, Sfi1p, localizes to the half...

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Bibliographic Details
Published in:The Journal of cell biology 2003-09, Vol.162 (7), p.1211-1221
Main Author: Kilmartin, John V.
Format: Article
Language:English
Subjects:
Alleles
Amino Acid Sequence
Amino acids
Binding Sites - physiology
Calcium - metabolism
Calcium-Binding Proteins - genetics
Calcium-Binding Proteins - metabolism
Cell cycle
Cell Cycle Proteins - genetics
Cell Cycle Proteins - metabolism
Cells
Cellular biology
Elasticity
Gels
Green Fluorescent Proteins
Humans
Indicators and Reagents - metabolism
Luminescent Proteins - genetics
Mitotic spindle apparatus
Molecular Sequence Data
Phenotype
Plasmids
Proteins
Repressor Proteins - chemistry
Repressor Proteins - genetics
Repressor Proteins - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Schizosaccharomyces
Schizosaccharomyces pombe Proteins
Sequence Homology, Amino Acid
Spindle Apparatus - metabolism
Spindle pole body
Yeast
Yeasts
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Summary:Centrins are calmodulin-like proteins present in microtubule-organizing centers. The Saccharomyces cerevisiae centrin, Cdc31p, was functionally tagged with a single Z domain of protein A, and used in pull-down experiments to isolate Cdc31p-binding proteins. One of these, Sfi1p, localizes to the half-bridge of the spindle pole body (SPB), where Cdc31p is also localized. Temperature-sensitive mutants in SFI1 show a defect in SPB duplication and genetic interactions with cdc31-1. Sfi1p contains multiple internal repeats that are also present in a Schizosaccharomyces pombe protein, which also localizes to the SPB, and in several human proteins, one of which localizes close to the centriole region. Cdc31p binds directly to individual Sfi1 repeats in a 1:1 ratio, so a single molecule of Sfi1p binds multiple molecules of Cdc31p. The centrosomal human protein containing Sfi1 repeats also binds centrin in the repeat region, showing that this centrin-binding motif is conserved.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.200307064