Mutational analysis of centromere DNA from chromosome VI of Saccharomyces cerevisiae

Journal Article · Wed Jun 01 00:00:00 EDT 1988 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.8.6.2523· OSTI ID:7002084

Hegemann, J H; Shero, J H; Cottarel, G; Philippsen, P; Hieter, P

Saccharomyces cerevisiae centromeres have a characteristic 120-base-pair region consisting of three different centromere DNA sequence elements (CDEI, CDEII, and CDEIII). The authors generated a series of 26 CEN mutations in vitro (including 22 point mutations, 3 insertions and 1 deletion) and tested their effects on mitotic chromosome segregation by using a new vector system. The yeast transformation vector pYCF5 was constructed to introduce wild-type mutant CEN DNAs onto large, linear chromosome fragments which are mitotically stable and nonessential. Six point mutations in CDEI show increased rates of chromosome loss events per cell division of 2- to 10-fold. Twenty mutations in CDEIII exhibit chromosome loss rates that vary from wild type (10/sup -4/) to nonfunctional (>10/sup -1/). These results directly identify nucleotides within CDEI and CDEIII that are required for the specification of a functional centromere and show that the degree of conservation of an individual base does not necessarily reflect its importance to mitotic CEN function.

• Research Organization: Dept. of Microbiology, Biocenter, Univ. of Basel, 4056 Basel (CH)
• OSTI ID: 7002084
• Journal Information: Mol. Cell. Biol.; (United States), Vol. 8:6
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
09 BIOMASS FUELS
CHROMOSOMAL ABERRATIONS
BIOLOGICAL PATHWAYS
CHROMOSOMES
STRUCTURE-ACTIVITY RELATIONSHIPS
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GENETIC MAPPING
DNA SEQUENCING
GENE MUTATIONS
MICROTUBULES
MITOSIS
MUTANTS
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CELL DIVISION
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