Genetic analysis of the mitotic transmission of minichromosomes

Douglas Koshland; John C. Kent; Leland H. Hartwell

doi:10.1016/0092-8674(85)90153-9

Genetic analysis of the mitotic transmission of minichromosomes

Douglas Koshland, John C. Kent, Leland H. Hartwell

Research output: Contribution to journal › Article › peer-review

232 Scopus citations

Abstract

The fidelity of the mitotic transmission of minichromosomes in S. cerevisiae is monitored by a novel visual assay that allows one to detect changes in plasmid copy number in individual mitotic divisions. This assay is used to investigate the mitotic transmission of a plasmid containing a putative yeast origin of replication (ARS1) and a centromere (CEN3). The rate of improper segregation for the minichromosome is 200-fold higher than observed for a normal chromosome. However, the replication of the minichromosome is stringently controlled; it overreplicates less than once per one thousand mitotic divisions. We also use this assay to isolate and characterize mutations in ARS1 and CEN3. The mutations in ARS1 define a new domain required for its optimal activity, and the mutations in CEN3 suggest that the integrity of element II is not essential for centromere function. Finally, the phenotypes of the mutations in ARS1 and CEN3 are consistent with their function in replication and segregation, respectively.

Original language	English (US)
Pages (from-to)	393-403
Number of pages	11
Journal	Cell
Volume	40
Issue number	2
DOIs	https://doi.org/10.1016/0092-8674(85)90153-9
State	Published - Feb 1985
Externally published	Yes

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/0092-8674(85)90153-9

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@article{d3788fb61d7d4592a41c2ecf6a121bac,

title = "Genetic analysis of the mitotic transmission of minichromosomes",

abstract = "The fidelity of the mitotic transmission of minichromosomes in S. cerevisiae is monitored by a novel visual assay that allows one to detect changes in plasmid copy number in individual mitotic divisions. This assay is used to investigate the mitotic transmission of a plasmid containing a putative yeast origin of replication (ARS1) and a centromere (CEN3). The rate of improper segregation for the minichromosome is 200-fold higher than observed for a normal chromosome. However, the replication of the minichromosome is stringently controlled; it overreplicates less than once per one thousand mitotic divisions. We also use this assay to isolate and characterize mutations in ARS1 and CEN3. The mutations in ARS1 define a new domain required for its optimal activity, and the mutations in CEN3 suggest that the integrity of element II is not essential for centromere function. Finally, the phenotypes of the mutations in ARS1 and CEN3 are consistent with their function in replication and segregation, respectively.",

author = "Douglas Koshland and Kent, {John C.} and Hartwell, {Leland H.}",

note = "Funding Information: We thank Steve Henikoff for the clone of the ADf3 gene. We are indebted to David Shortle for valuable suggestions for performing the in vitro mutagenesis of pDK206 and to Judith Campbell, John Carbon, and R. McCarroll for results prior to publication. We thank Robert McCarroll for measuring the simple loss rate of a plasmid containing two ARS sequences. We are grateful to Bonita Brewer, Dan Burke, Michael Carson, Walt Fangman, Nancy Hollingsworth, and Deborah Kranzler for critical reading of the manuscript. D. K. is a fellow of the Helen Hay Whitney Foundation. Research was supported by grants",

year = "1985",

month = feb,

doi = "10.1016/0092-8674(85)90153-9",

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pages = "393--403",

journal = "Cell",

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T1 - Genetic analysis of the mitotic transmission of minichromosomes

AU - Koshland, Douglas

AU - Kent, John C.

AU - Hartwell, Leland H.

N1 - Funding Information: We thank Steve Henikoff for the clone of the ADf3 gene. We are indebted to David Shortle for valuable suggestions for performing the in vitro mutagenesis of pDK206 and to Judith Campbell, John Carbon, and R. McCarroll for results prior to publication. We thank Robert McCarroll for measuring the simple loss rate of a plasmid containing two ARS sequences. We are grateful to Bonita Brewer, Dan Burke, Michael Carson, Walt Fangman, Nancy Hollingsworth, and Deborah Kranzler for critical reading of the manuscript. D. K. is a fellow of the Helen Hay Whitney Foundation. Research was supported by grants

PY - 1985/2

Y1 - 1985/2

N2 - The fidelity of the mitotic transmission of minichromosomes in S. cerevisiae is monitored by a novel visual assay that allows one to detect changes in plasmid copy number in individual mitotic divisions. This assay is used to investigate the mitotic transmission of a plasmid containing a putative yeast origin of replication (ARS1) and a centromere (CEN3). The rate of improper segregation for the minichromosome is 200-fold higher than observed for a normal chromosome. However, the replication of the minichromosome is stringently controlled; it overreplicates less than once per one thousand mitotic divisions. We also use this assay to isolate and characterize mutations in ARS1 and CEN3. The mutations in ARS1 define a new domain required for its optimal activity, and the mutations in CEN3 suggest that the integrity of element II is not essential for centromere function. Finally, the phenotypes of the mutations in ARS1 and CEN3 are consistent with their function in replication and segregation, respectively.

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Genetic analysis of the mitotic transmission of minichromosomes

Abstract

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