The objectives are to define the structural organization of the centromere regions of eukaryotic chromosomes, and to understand the molecular mechanisms involved in chromosome segregation during mitotic and meiotic cell divisions. The centromere studies supported by this grant are carried out using as experimental systems the budding yeast, Saccharomyces cerevisiae, and the filamentous fungus, Aspergillus nidulans. Synthetic minichromosomes, containing a functional centromere (CEN), a DNA replicator function, and telomeric DNA (if linear), constitute invaluable probes for the study of chromosome structure and function.
Specific aims i nclude: (1) the construction of structurally altered centromere DNAs with altered function, using in vitro mutagenesis techniques; (2) the identification of genes specifying kinetochore structural proteins or gene products necessary for formation of the yeast kinetochore; (3) the isolation and characterization of kinetochore proteins (centromere binding proteins); (4) investigation of the mechanism of kinetochore-microtubule associations; (5) investigation of specialized DNA structures in the centromere region, including replicative intermediates, kinked DNA, and methylated DNA; (6) isolation and structural characterization of centromere DNAs from Aspergillus nidulans, as a step toward an understanding of centromere structure-function in more complex organisms. Studies on the molecular mechanisms involved in cell division and chromosome segregation in the experimentally tractable lower eukaryotes offer an approach to an understanding of analogous mechanisms as they function normally and abnormally in higher organisms.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA011034-20
Application #
3163426
Study Section
Genetics Study Section (GEN)
Project Start
1977-01-01
Project End
1991-12-31
Budget Start
1988-01-01
Budget End
1988-12-31
Support Year
20
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of California Santa Barbara
Department
Type
Schools of Arts and Sciences
DUNS #
City
Santa Barbara
State
CA
Country
United States
Zip Code
93106
Mishra, Prashant K; Baum, Mary; Carbon, John (2011) DNA methylation regulates phenotype-dependent transcriptional activity in Candida albicans. Proc Natl Acad Sci U S A 108:11965-70
Mishra, Prashant K; Baum, Mary; Carbon, John (2007) Centromere size and position in Candida albicans are evolutionarily conserved independent of DNA sequence heterogeneity. Mol Genet Genomics 278:455-65
Baum, Mary; Sanyal, Kaustuv; Mishra, Prashant K et al. (2006) Formation of functional centromeric chromatin is specified epigenetically in Candida albicans. Proc Natl Acad Sci U S A 103:14877-82
Sanyal, Kaustuv; Baum, Mary; Carbon, John (2004) Centromeric DNA sequences in the pathogenic yeast Candida albicans are all different and unique. Proc Natl Acad Sci U S A 101:11374-9
Stoyan, Tanja; Carbon, John (2004) Inner kinetochore of the pathogenic yeast Candida glabrata. Eukaryot Cell 3:1154-63
Yoon, H-J (2004) A novel small-molecule inhibitor of the chromosome segregation process in yeast. Mol Genet Genomics 271:490-8
Sanyal, Kaustuv; Carbon, John (2002) The CENP-A homolog CaCse4p in the pathogenic yeast Candida albicans is a centromere protein essential for chromosome transmission. Proc Natl Acad Sci U S A 99:12969-74
Stoyan, T; Gloeckner, G; Diekmann, S et al. (2001) Multifunctional centromere binding factor 1 is essential for chromosome segregation in the human pathogenic yeast Candida glabrata. Mol Cell Biol 21:4875-88
Pietrasanta, L I; Thrower, D; Hsieh, W et al. (1999) Probing the Saccharomyces cerevisiae centromeric DNA (CEN DNA)-binding factor 3 (CBF3) kinetochore complex by using atomic force microscopy. Proc Natl Acad Sci U S A 96:3757-62
Zebarjadian, Y; King, T; Fournier, M J et al. (1999) Point mutations in yeast CBF5 can abolish in vivo pseudouridylation of rRNA. Mol Cell Biol 19:7461-72

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