Two genes required for the binding of an essential Saccharomyces cerevisiae kinetochore complex to DNA

• Published in: Proceedings of the National Academy of Sciences - PNAS 1995-12, Vol.92 (26), p.12026-12030
• Main Authors: Sorger, P K, Doheny, K F, Hieter, P, Kopski, K M, Huffaker, T C, Hyman, A A
• Format: Article
• Language: English
• Subjects: adn; Base Sequence; binding proteins; Cell growth; Cell separation; Cells; Cellular biology; Centromere - physiology; Centromeres; chromosome; Chromosomes; Chromosomes, Fungal; cromosomas; Deoxyribonucleic acid; DNA; DNA Footprinting; DNA probes; DNA, Fungal - chemistry; DNA, Fungal - genetics; DNA, Fungal - metabolism; DNA-Binding Proteins - metabolism; Fungal Proteins - metabolism; Genes; Genes, Fungal; Genetic mutation; Kinetochores; Kinetochores - physiology; Macromolecular Substances; Molecular biology; Molecular Sequence Data; mutacion; Mutagenesis, Site-Directed; mutant; mutantes; mutants; mutation; Nuclear Proteins; nucleotide sequence; Oligodeoxyribonucleotides; proteinas aglutinantes; proteine de liaison; Recombinant Proteins - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins; secuencia nucleica; sequence nucleique; Yeast
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.26.12026

Kinetochores are DNA-protein structures that assemble on centromeric DNA and attach chromosomes to spindle microtubules. Because of their simplicity, the 125-bp centromeres of Saccharomyces cerevisiae are particularly amenable to molecular analysis. Budding yeast centromeres contain three sequence elements of which centromere DNA sequence element III (CDEIII) appears to be particularly important. cis-acting mutations in CDEIII and trans-acting mutations in genes encoding subunits of the CDEIII-binding complex (CBF3) prevent correct chromosome transmission. Using temperature-sensitive mutations in CBF3 subunits, we show a strong correlation between DNA-binding activity measured in vivo and kinetochore activity in vivo. We extend previous findings by Goh and Kilmartin [Goh, P.-Y. and Kilmartin, J. V. (1993) J. Cell Biol. 121, 503-512] to argue that DNA-bound CBF3 may be involved in the operation of a mitotic checkpoint but that functional CBF3 is not required for the assembly of a bipolar spindle.