Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria

Sarah L. Donahue; Uma Lakshmipathy; Colin R Campbell

doi:10.1016/S1567-7249(02)00037-5

Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria

Sarah L. Donahue, Uma Lakshmipathy, Colin R Campbell

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

1 Scopus citations

Abstract

Expression vectors were created in which the 5′ end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.

Original language	English (US)
Pages (from-to)	47-57
Number of pages	11
Journal	Mitochondrion
Volume	2
Issue number	1-2
DOIs	https://doi.org/10.1016/S1567-7249(02)00037-5
State	Published - Nov 2002

Bibliographical note

Funding Information:
We thank Drs Barbara Garvik and Lee Hartwell as well as Brendan Cormack, Jasper Rine, and Masimo Zeviani for kindly providing yeast strains and reagents used herein. We also thank Dr Modrich for kindly providing Eco R I antibody. This work was supported in part by a grant from the NIH (AG16678) and by a grant from the American Heart Association (AHA MN 9951198Z).

Keywords

Mitochondria
Restriction endonuclease
Saccharomyces cerevisiae
Targeting

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10.1016/S1567-7249(02)00037-5

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title = "Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria",

abstract = "Expression vectors were created in which the 5′ end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.",

keywords = "Mitochondria, Restriction endonuclease, Saccharomyces cerevisiae, Targeting",

author = "Donahue, {Sarah L.} and Uma Lakshmipathy and Campbell, {Colin R}",

note = "Funding Information: We thank Drs Barbara Garvik and Lee Hartwell as well as Brendan Cormack, Jasper Rine, and Masimo Zeviani for kindly providing yeast strains and reagents used herein. We also thank Dr Modrich for kindly providing Eco R I antibody. This work was supported in part by a grant from the NIH (AG16678) and by a grant from the American Heart Association (AHA MN 9951198Z).",

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doi = "10.1016/S1567-7249(02)00037-5",

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T1 - Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria

AU - Donahue, Sarah L.

AU - Lakshmipathy, Uma

AU - Campbell, Colin R

N1 - Funding Information: We thank Drs Barbara Garvik and Lee Hartwell as well as Brendan Cormack, Jasper Rine, and Masimo Zeviani for kindly providing yeast strains and reagents used herein. We also thank Dr Modrich for kindly providing Eco R I antibody. This work was supported in part by a grant from the NIH (AG16678) and by a grant from the American Heart Association (AHA MN 9951198Z).

PY - 2002/11

Y1 - 2002/11

N2 - Expression vectors were created in which the 5′ end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.

AB - Expression vectors were created in which the 5′ end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.

KW - Mitochondria

KW - Restriction endonuclease

KW - Saccharomyces cerevisiae

KW - Targeting

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SN - 1567-7249

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JO - Mitochondrion

JF - Mitochondrion

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ER -

Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria

Abstract

Bibliographical note

Keywords

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