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CEP3 encodes a centromere protein of Saccharomyces cerevisiae
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...
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| Published in: | The Journal of cell biology 1995-03, Vol.128 (5), p.749-760 |
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| container_issue | 5 |
| container_start_page | 749 |
| container_title | The Journal of cell biology |
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| creator | Strunnikov, A.V. (Carnegie Institution of Washington, Baltimore, Maryland) Kingsbury, J Koshland, D |
| description | 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) |
| doi_str_mv | 10.1083/jcb.128.5.749 |
| format | article |
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(Carnegie Institution of Washington, Baltimore, Maryland) ; Kingsbury, J ; Koshland, D</creator><creatorcontrib>Strunnikov, A.V. (Carnegie Institution of Washington, Baltimore, Maryland) ; Kingsbury, J ; Koshland, D</creatorcontrib><description>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)</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.128.5.749</identifier><identifier>PMID: 7876302</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>United States: Rockefeller University Press</publisher><subject>ADN ; Amino Acid Sequence ; Base Sequence ; Cells ; Cellular biology ; Centromere - genetics ; Centromere - physiology ; Centromeres ; CHROMOSOME ; Chromosomes ; Chromosomes, Fungal - genetics ; Cloning, Molecular ; COMPOSICION QUIMICA ; COMPOSITION CHIMIQUE ; CROMOSOMAS ; Daughter cells ; DNA ; DNA-Binding Proteins - genetics ; ESTRUCTURA CELULAR ; GENE ; GENES ; Genes, Fungal - genetics ; Genes, Lethal - genetics ; Genetic mutation ; Kinetochores ; Microtubules - physiology ; Models, Genetic ; Molecular Sequence Data ; Mutagenesis ; MUTANT ; MUTANTES ; Mutation ; Nuclear Proteins - genetics ; Plasmids ; PROTEINAS ; PROTEINE ; Restriction Mapping ; SACCHAROMYCES CEREVISIAE ; Saccharomyces cerevisiae - genetics ; Saccharomyces cerevisiae - ultrastructure ; Saccharomyces cerevisiae Proteins ; SECUENCIA NUCLEICA ; SEGREGACION ; SEGREGATION ; Selection, Genetic ; Sequence Analysis, DNA ; SEQUENCE NUCLEIQUE ; Spindle Apparatus - physiology ; STRUCTURE CELLULAIRE ; Yeast ; Yeasts</subject><ispartof>The Journal of cell biology, 1995-03, Vol.128 (5), p.749-760</ispartof><rights>Copyright 1995 The Rockefeller University Press</rights><rights>Copyright Rockefeller University Press Mar 1995</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c480t-ecd2aece1a752ef46b354072b2cfe0a599f7e7cd7df628e6b447aa801db236953</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7876302$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Strunnikov, A.V. (Carnegie Institution of Washington, Baltimore, Maryland)</creatorcontrib><creatorcontrib>Kingsbury, J</creatorcontrib><creatorcontrib>Koshland, D</creatorcontrib><title>CEP3 encodes a centromere protein of Saccharomyces cerevisiae</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>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)</description><subject>ADN</subject><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Cells</subject><subject>Cellular biology</subject><subject>Centromere - genetics</subject><subject>Centromere - physiology</subject><subject>Centromeres</subject><subject>CHROMOSOME</subject><subject>Chromosomes</subject><subject>Chromosomes, Fungal - genetics</subject><subject>Cloning, Molecular</subject><subject>COMPOSICION QUIMICA</subject><subject>COMPOSITION CHIMIQUE</subject><subject>CROMOSOMAS</subject><subject>Daughter cells</subject><subject>DNA</subject><subject>DNA-Binding Proteins - genetics</subject><subject>ESTRUCTURA CELULAR</subject><subject>GENE</subject><subject>GENES</subject><subject>Genes, Fungal - genetics</subject><subject>Genes, Lethal - genetics</subject><subject>Genetic mutation</subject><subject>Kinetochores</subject><subject>Microtubules - physiology</subject><subject>Models, Genetic</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis</subject><subject>MUTANT</subject><subject>MUTANTES</subject><subject>Mutation</subject><subject>Nuclear Proteins - genetics</subject><subject>Plasmids</subject><subject>PROTEINAS</subject><subject>PROTEINE</subject><subject>Restriction Mapping</subject><subject>SACCHAROMYCES CEREVISIAE</subject><subject>Saccharomyces cerevisiae - genetics</subject><subject>Saccharomyces cerevisiae - ultrastructure</subject><subject>Saccharomyces cerevisiae Proteins</subject><subject>SECUENCIA NUCLEICA</subject><subject>SEGREGACION</subject><subject>SEGREGATION</subject><subject>Selection, Genetic</subject><subject>Sequence Analysis, DNA</subject><subject>SEQUENCE NUCLEIQUE</subject><subject>Spindle Apparatus - physiology</subject><subject>STRUCTURE CELLULAIRE</subject><subject>Yeast</subject><subject>Yeasts</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><recordid>eNpdkUtLAzEUhYMoWqtLN6IwuHA39eY9s1CQ4gsEBXUdMpk7OqWd1GQq-O-NtvhaBfJ9HE5yCNmjMKJQ8JOJq0aUFSM50qJcIwMqBeQFFbBOBgCM5qVkcotsxzgBAKEF3ySbutCKAxuQ0_HFPc-wc77GmNnMYdcHP8OA2Tz4Htsu8032YJ17sen-3SXLJfrWxtbiDtlo7DTi7uockqfLi8fxdX57d3UzPr_NnSigz9HVzKJDarVk2AhV8VRSs4q5BsHKsmw0alfrulGsQFUJoa0tgNYV46qUfEjOlrnzRTXD-quknZp5aGc2vBtvW_OXdO2LefZvhlEGvKQp4HgVEPzrAmNvZm10OJ3aDv0iGqq0KkTJknj0T5z4RejS41KWpqAh_fmQ5EvJBR9jwOa7CQXzOYpJo5g0ipEmjZL8w9_1v-3VCokfLPkk9j78hCmqlOIJ7y9xY72xz6GN5umhlIJTqfkHnn-Z8g</recordid><startdate>19950301</startdate><enddate>19950301</enddate><creator>Strunnikov, A.V. 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(Carnegie Institution of Washington, Baltimore, Maryland) ; Kingsbury, J ; Koshland, D</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c480t-ecd2aece1a752ef46b354072b2cfe0a599f7e7cd7df628e6b447aa801db236953</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>ADN</topic><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Cells</topic><topic>Cellular biology</topic><topic>Centromere - genetics</topic><topic>Centromere - physiology</topic><topic>Centromeres</topic><topic>CHROMOSOME</topic><topic>Chromosomes</topic><topic>Chromosomes, Fungal - genetics</topic><topic>Cloning, Molecular</topic><topic>COMPOSICION QUIMICA</topic><topic>COMPOSITION CHIMIQUE</topic><topic>CROMOSOMAS</topic><topic>Daughter cells</topic><topic>DNA</topic><topic>DNA-Binding Proteins - genetics</topic><topic>ESTRUCTURA CELULAR</topic><topic>GENE</topic><topic>GENES</topic><topic>Genes, Fungal - genetics</topic><topic>Genes, Lethal - genetics</topic><topic>Genetic mutation</topic><topic>Kinetochores</topic><topic>Microtubules - physiology</topic><topic>Models, Genetic</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis</topic><topic>MUTANT</topic><topic>MUTANTES</topic><topic>Mutation</topic><topic>Nuclear Proteins - genetics</topic><topic>Plasmids</topic><topic>PROTEINAS</topic><topic>PROTEINE</topic><topic>Restriction Mapping</topic><topic>SACCHAROMYCES CEREVISIAE</topic><topic>Saccharomyces cerevisiae - genetics</topic><topic>Saccharomyces cerevisiae - ultrastructure</topic><topic>Saccharomyces cerevisiae Proteins</topic><topic>SECUENCIA NUCLEICA</topic><topic>SEGREGACION</topic><topic>SEGREGATION</topic><topic>Selection, Genetic</topic><topic>Sequence Analysis, DNA</topic><topic>SEQUENCE NUCLEIQUE</topic><topic>Spindle Apparatus - physiology</topic><topic>STRUCTURE CELLULAIRE</topic><topic>Yeast</topic><topic>Yeasts</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Strunnikov, A.V. 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(Carnegie Institution of Washington, Baltimore, Maryland)</au><au>Kingsbury, J</au><au>Koshland, D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>CEP3 encodes a centromere protein of Saccharomyces cerevisiae</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1995-03-01</date><risdate>1995</risdate><volume>128</volume><issue>5</issue><spage>749</spage><epage>760</epage><pages>749-760</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><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)</abstract><cop>United States</cop><pub>Rockefeller University Press</pub><pmid>7876302</pmid><doi>10.1083/jcb.128.5.749</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of cell biology, 1995-03, Vol.128 (5), p.749-760 |
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| source | Alma/SFX Local Collection |
| subjects | ADN Amino Acid Sequence Base Sequence Cells Cellular biology Centromere - genetics Centromere - physiology Centromeres CHROMOSOME Chromosomes Chromosomes, Fungal - genetics Cloning, Molecular COMPOSICION QUIMICA COMPOSITION CHIMIQUE CROMOSOMAS Daughter cells DNA DNA-Binding Proteins - genetics ESTRUCTURA CELULAR GENE GENES Genes, Fungal - genetics Genes, Lethal - genetics Genetic mutation Kinetochores Microtubules - physiology Models, Genetic Molecular Sequence Data Mutagenesis MUTANT MUTANTES Mutation Nuclear Proteins - genetics Plasmids PROTEINAS PROTEINE Restriction Mapping SACCHAROMYCES CEREVISIAE Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - ultrastructure Saccharomyces cerevisiae Proteins SECUENCIA NUCLEICA SEGREGACION SEGREGATION Selection, Genetic Sequence Analysis, DNA SEQUENCE NUCLEIQUE Spindle Apparatus - physiology STRUCTURE CELLULAIRE Yeast Yeasts |
| title | CEP3 encodes a centromere protein of Saccharomyces cerevisiae |
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