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PMID: 14504268 Published · ppublish English Journal Article

Sfi1p has conserved centrin-binding sites and an essential function in budding yeast spindle pole body duplication.

The Journal of cell biology ·Vol. 162 ·No. 7 ·2003-09-29 ·Pages 1211-21

Kilmartin JV

Abstract

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.

MeSH Terms
Amino Acid Sequence Binding Sites/physiology Calcium/metabolism Calcium-Binding Proteins/genetics,metabolism Cell Cycle Proteins/genetics,metabolism Elasticity Green Fluorescent Proteins Humans Indicators and Reagents/metabolism Luminescent Proteins/genetics Molecular Sequence Data Phenotype Repressor Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Schizosaccharomyces Schizosaccharomyces pombe Proteins Sequence Homology, Amino Acid Spindle Apparatus/metabolism
Chemicals
CDC31 protein, S cerevisiae Calcium-Binding Proteins Cell Cycle Proteins Indicators and Reagents Luminescent Proteins Repressor Proteins SFI1 protein, S cerevisiae Saccharomyces cerevisiae Proteins Schizosaccharomyces pombe Proteins cdc31 protein, S pombe Green Fluorescent Proteins Calcium
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Kilmartin John V
MRC Laboratory of Molecular Biology, Cambridge CB2 2QH, UK. jvk@mrc-lmb.cam.ac.uk
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200307064
Email
jvk@mrc-lmb.cam.ac.uk
Published
2003-09-29
Epub
2003-00-22
Pages
1211-21
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2173958
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.200307064 PMC Full Text View journal details
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