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PMID: 7876302 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

CEP3 encodes a centromere protein of Saccharomyces cerevisiae.

The Journal of cell biology ·Vol. 128 ·No. 5 ·1995-03-00 ·Pages 749-60

Strunnikov AV, Kingsbury J, Koshland D

Abstract

We have designed a screen to identify mutants specifically affecting kinetochore function in the yeast Saccharomyces cerevisiae. The selection procedure was based on the generation of "synthetic acentric" minichromosomes. "Synthetic acentric" minichromosomes contain a centromere locus, but lack centromere activity due to combination of mutations in centromere DNA and in a chromosomal gene (CEP) encoding a putative centromere protein. Ten conditional lethal cep mutants were isolated, seven were found to be alleles of NDC10 (CEP2) encoding the 110-kD protein of yeast kinetochore. Three mutants defined a novel essential gene CEP3. The CEP3 product (Cep3p) is a 71-kD protein with a potential DNA-binding domain (binuclear Zn-cluster). At nonpermissive temperature the cep3 cells arrest with an undivided nucleus and a short mitotic spindle. At permissive temperature the cep3 cells are unable to support segregation of minichromosomes with mutations in the central part of element III of yeast centromere DNA. These minichromosomes, when isolated from cep3 cultures, fail to bind bovine microtubules in vitro. The sum of genetic, cytological and biochemical data lead us to suggest that the Cep3 protein is a DNA-binding component of yeast centromere. Molecular mass and sequence comparison confirm that Cep3p is the p64 component of centromere DNA binding complex Cbf3 (Lechner, 1994).

Related Genes
CEP3
MeSH Terms
Amino Acid Sequence Base Sequence Centromere/genetics,physiology Chromosomes, Fungal/genetics Cloning, Molecular DNA-Binding Proteins/genetics Genes, Fungal/genetics Genes, Lethal/genetics Kinetochores Microtubules/physiology Models, Genetic Molecular Sequence Data Mutagenesis Nuclear Proteins/genetics Restriction Mapping Saccharomyces cerevisiae/genetics,ultrastructure Saccharomyces cerevisiae Proteins Selection, Genetic Sequence Analysis, DNA Spindle Apparatus/physiology
Chemicals
CEP3 protein, S cerevisiae DNA-Binding Proteins Nuclear Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Strunnikov A V
Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210.
Kingsbury J
Koshland D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.128.5.749
Published
1995-03-00
Pages
749-60
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120391
Subset
IM
Grants
NIGMS NIH HHS · GM-41718 · United States
Databases
GENBANK
U12339, X79803
External Links
PubMed source DOI: 10.1083/jcb.128.5.749 PMC Full Text View journal details
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