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Chromosoma

, Volume 101, Issue 3, pp 189–197 | Cite as

Analysis of centromere function in Saccharomyces cerevisiae using synthetic centromere mutants

  • Michael R. Murphy
  • Dana M. Fowlkes
  • Molly Fitzgerald-Hayes
Original Articles

Abstract

We constructed Saccharomyces cerevisiae centromere DNA mutants by annealing and ligating synthetic oligonucleotides, a novel approach to centromere DNA mutagenesis that allowed us to change only one structural parameter at a time. Using this method, we confirmed that CDE I, II, and III alone are sufficient for centromere function and that A+T-rich sequences in CDE II play important roles in mitosis and meiosis. Analysis of mutants also showed that a bend in the centromere DNA could be important for proper mitotic and meiotic chromosome segregation. In addition we demonstrated that the wild-type orientation of the CDE III sequence, but not the CDE I sequence, is critical for wild-type mitotic segregation. Surprisingly, we found that one mutant centromere affected the segregation of plasmids and chromosomes differently. The implications of these results for centromere function and chromosome structure are discussed.

Keywords

Developmental Biology Saccharomyces Cerevisiae Chromosome Segregation Chromosome Structure Meiotic Chromosome 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Michael R. Murphy
    • 1
  • Dana M. Fowlkes
    • 2
  • Molly Fitzgerald-Hayes
    • 1
  1. 1.Department of Biochemistry, Program in Molecular and Cellular Biology, 901 Lederle Graduate Research CenterUniversity of Massachusetts at AmherstAmherstUSA
  2. 2.Department of Pathology, Curriculum in GeneticsUniversity of North Carolina at Chapel HillChapel HillUSA

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