Centromere DNA from yeast can be isolated, inserted into plasmids, and shown to exhibit the basic behavioral characteristics of intact centromeres in the genome. In these plasmids, centromere structure-function relationships are especially amenable to direct study. We will use this approach in the proposed research to investigate the molecular details of processes by which the centomere governs chromosome organization and movement in yeast. In previous work we have shown that two yeast centromere DNAs (CEN3 and CEN11) contain conserved arrangements of DNA sequence homology which are essential for centromere function. We will extend these observations by isolating and characterizing CEN7 from a very large yeast chromosome (VII), to examine structural similarities and differences. Using in vitro mutagenesis techniques to alter centromere plasmids, we will analyze the effect of specific sequence changes and structural rearrangements on centromere function. We will also study centromeric-flanking DNA segments that, as indicated by preliminary studies, may function to assure proper chromosome segregation and to maintain specific chromatin configurations. In addition, we will investigate a possible role for DNA methylation in centromere function. Yeast centromere DNA sequences are organized in a unique, nuclease-resistant chromatin core which may function as a primitive kinetochore. The single microtubule attachment site in yeast may, in fact, be a model repeat unit structure for the multiple attachment sites found in other, more complex, kinetochores.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032257-03
Application #
3280930
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1983-07-01
Project End
1986-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Massachusetts Amherst
Department
Type
Schools of Arts and Sciences
DUNS #
153223151
City
Amherst
State
MA
Country
United States
Zip Code
Schroeder, A J; Chen, X H; Xiao, Z et al. (1999) Genetic evidence for interactions between yeast importin alpha (Srp1p) and its nuclear export receptor, Cse1p. Mol Gen Genet 261:788-95
Xiao, Z X; Fitzgerald-Hayes, M (1995) Functional interaction between the CSE2 gene product and centromeres in Saccharomyces cerevisiae. J Mol Biol 248:255-63
Stoler, S; Keith, K C; Curnick, K E et al. (1995) A mutation in CSE4, an essential gene encoding a novel chromatin-associated protein in yeast, causes chromosome nondisjunction and cell cycle arrest at mitosis. Genes Dev 9:573-86
Chen, X H; Xiao, Z; Fitzgerald-Hayes, M (1994) SCM2, a tryptophan permease in Saccharomyces cerevisiae, is important for cell growth. Mol Gen Genet 244:260-8
Xiao, Z; McGrew, J T; Schroeder, A J et al. (1993) CSE1 and CSE2, two new genes required for accurate mitotic chromosome segregation in Saccharomyces cerevisiae. Mol Cell Biol 13:4691-702
Payne, W E; Fitzgerald-Hayes, M (1993) A mutation in PLC1, a candidate phosphoinositide-specific phospholipase C gene from Saccharomyces cerevisiae, causes aberrant mitotic chromosome segregation. Mol Cell Biol 13:4351-64
Murphy, M R; Fowlkes, D M; Fitzgerald-Hayes, M (1991) Analysis of centromere function in Saccharomyces cerevisiae using synthetic centromere mutants. Chromosoma 101:189-97
Densmore, L; Payne, W E; Fitzgerald-Hayes, M (1991) In vivo genomic footprint of a yeast centromere. Mol Cell Biol 11:154-65
Murphy, M; Fitzgerald-Hayes, M (1990) Cis- and trans-acting factors involved in centromere function in Saccharomyces cerevisiae. Mol Microbiol 4:329-36
Gaudet, A; Fitzgerald-Hayes, M (1989) Mutations in CEN3 cause aberrant chromosome segregation during meiosis in Saccharomyces cerevisiae. Genetics 121:477-89

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