Mutational analysis of centromere DNA from chromosome VI of Saccharomyces cerevisiae - PubMed (original) (raw)

Mutational analysis of centromere DNA from chromosome VI of Saccharomyces cerevisiae

J H Hegemann et al. Mol Cell Biol. 1988 Jun.

Abstract

Saccharomyces cerevisiae centromeres have a characteristic 120-base-pair region consisting of three distinct centromere DNA sequence elements (CDEI, CDEII, and CDEIII). We have 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 and 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(-4)) to nonfunctional (greater than 10(-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 in mitotic CEN function.

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References

    1. Cell. 1984 May;37(1):67-75 - PubMed
    1. Gene. 1983 Dec;26(1):101-6 - PubMed
    1. Nucleic Acids Res. 1984 Jul 25;12(14):5647-64 - PubMed
    1. Annu Rev Biochem. 1984;53:293-321 - PubMed
    1. Cell. 1984 Nov;39(1):191-201 - PubMed

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