Intramitochondrial transfer and engineering of mammalian mitochondrial genomes in yeast

Young Geol Yoon; Michael D. Koob

doi:10.1016/j.mito.2019.03.006

Intramitochondrial transfer and engineering of mammalian mitochondrial genomes in yeast

Young Geol Yoon, Michael D. Koob

Laboratory Medicine and Pathology

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

5 Scopus citations

Abstract

Mitochondrial genomes (mtDNA) depend on the nuclear genome with which they have evolved to provide essential replication functions and have been known to replicate as xenotransplants only in the cells of closely related species. We now report that complete mouse mitochondrial genomes can be stably transplanted into the mitochondrial network in yeast devoid of their own mtDNA. Our analyses of these xenomitochondrial yeast cells show that they are accurately replicating intact mouse mtDNA genomes without rearrangement and that these mtDNA genomes have the same overall topology as the mtDNA present in the mouse mitochondrial network (i.e., circular monomers). Moreover, non-mtDNA replication and selection sequences required for maintaining the mitochondrial genomes in bacterial hosts are dispensable in these yeast mitochondria and could be efficiently and seamlessly removed by targeted homologous recombination within the mitochondria. These findings demonstrate that the yeast mtDNA replication system is capable of accurately replicating intact mammalian mtDNA genomes without sequence loss or rearrangement and that yeast mitochondria are a highly versatile host system for engineering complete mammalian mitochondrial genomes.

Original language	English (US)
Pages (from-to)	15-21
Number of pages	7
Journal	Mitochondrion
Volume	46
DOIs	https://doi.org/10.1016/j.mito.2019.03.006
State	Published - May 2019

Bibliographical note

Funding Information:
We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [R21NS052612, R21NS064398 to M.K.]; the Minnesota Medical Foundation; the Minnesota Partnership for Biotechnology and Medical Genomics; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [2011-0008052 and 2015M3A6A8066204 to Y.Y.].

Funding Information:
We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [ R21NS052612 , R21NS064398 to M.K.]; the Minnesota Medical Foundation ; the Minnesota Partnership for Biotechnology and Medical Genomics ; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [ 2011-0008052 and 2015M3A6A8066204 to Y.Y.].

Publisher Copyright:
© 2019 Elsevier B.V. and Mitochondria Research Society

Keywords

Mitochondrial genome
Mitochondrial host
Non-mtDNA sequence
Petite mutant
Xenomitochondrial yeast
mtDNA engineering

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10.1016/j.mito.2019.03.006

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title = "Intramitochondrial transfer and engineering of mammalian mitochondrial genomes in yeast",

abstract = "Mitochondrial genomes (mtDNA) depend on the nuclear genome with which they have evolved to provide essential replication functions and have been known to replicate as xenotransplants only in the cells of closely related species. We now report that complete mouse mitochondrial genomes can be stably transplanted into the mitochondrial network in yeast devoid of their own mtDNA. Our analyses of these xenomitochondrial yeast cells show that they are accurately replicating intact mouse mtDNA genomes without rearrangement and that these mtDNA genomes have the same overall topology as the mtDNA present in the mouse mitochondrial network (i.e., circular monomers). Moreover, non-mtDNA replication and selection sequences required for maintaining the mitochondrial genomes in bacterial hosts are dispensable in these yeast mitochondria and could be efficiently and seamlessly removed by targeted homologous recombination within the mitochondria. These findings demonstrate that the yeast mtDNA replication system is capable of accurately replicating intact mammalian mtDNA genomes without sequence loss or rearrangement and that yeast mitochondria are a highly versatile host system for engineering complete mammalian mitochondrial genomes.",

keywords = "Mitochondrial genome, Mitochondrial host, Non-mtDNA sequence, Petite mutant, Xenomitochondrial yeast, mtDNA engineering",

author = "Yoon, {Young Geol} and Koob, {Michael D.}",

note = "Funding Information: We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [R21NS052612, R21NS064398 to M.K.]; the Minnesota Medical Foundation; the Minnesota Partnership for Biotechnology and Medical Genomics; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [2011-0008052 and 2015M3A6A8066204 to Y.Y.]. Funding Information: We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [ R21NS052612 , R21NS064398 to M.K.]; the Minnesota Medical Foundation ; the Minnesota Partnership for Biotechnology and Medical Genomics ; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [ 2011-0008052 and 2015M3A6A8066204 to Y.Y.]. Publisher Copyright: {\textcopyright} 2019 Elsevier B.V. and Mitochondria Research Society",

year = "2019",

month = may,

doi = "10.1016/j.mito.2019.03.006",

language = "English (US)",

volume = "46",

pages = "15--21",

journal = "Mitochondrion",

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AU - Koob, Michael D.

N1 - Funding Information: We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [R21NS052612, R21NS064398 to M.K.]; the Minnesota Medical Foundation; the Minnesota Partnership for Biotechnology and Medical Genomics; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [2011-0008052 and 2015M3A6A8066204 to Y.Y.]. Funding Information: We would like to thank Drs. Thomas D. Fox, Dennis Livingstone and Kathleen Conklin for their gifts of plasmids and yeast strains. This work was supported by the National Institutes of Health [ R21NS052612 , R21NS064398 to M.K.]; the Minnesota Medical Foundation ; the Minnesota Partnership for Biotechnology and Medical Genomics ; the Muscular Dystrophy Association and the Friedreich's Ataxia Research Alliance [all to M.K.]; and the National Research Foundation of Korea Grant funded by the Korean government [ 2011-0008052 and 2015M3A6A8066204 to Y.Y.]. Publisher Copyright: © 2019 Elsevier B.V. and Mitochondria Research Society

PY - 2019/5

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N2 - Mitochondrial genomes (mtDNA) depend on the nuclear genome with which they have evolved to provide essential replication functions and have been known to replicate as xenotransplants only in the cells of closely related species. We now report that complete mouse mitochondrial genomes can be stably transplanted into the mitochondrial network in yeast devoid of their own mtDNA. Our analyses of these xenomitochondrial yeast cells show that they are accurately replicating intact mouse mtDNA genomes without rearrangement and that these mtDNA genomes have the same overall topology as the mtDNA present in the mouse mitochondrial network (i.e., circular monomers). Moreover, non-mtDNA replication and selection sequences required for maintaining the mitochondrial genomes in bacterial hosts are dispensable in these yeast mitochondria and could be efficiently and seamlessly removed by targeted homologous recombination within the mitochondria. These findings demonstrate that the yeast mtDNA replication system is capable of accurately replicating intact mammalian mtDNA genomes without sequence loss or rearrangement and that yeast mitochondria are a highly versatile host system for engineering complete mammalian mitochondrial genomes.

AB - Mitochondrial genomes (mtDNA) depend on the nuclear genome with which they have evolved to provide essential replication functions and have been known to replicate as xenotransplants only in the cells of closely related species. We now report that complete mouse mitochondrial genomes can be stably transplanted into the mitochondrial network in yeast devoid of their own mtDNA. Our analyses of these xenomitochondrial yeast cells show that they are accurately replicating intact mouse mtDNA genomes without rearrangement and that these mtDNA genomes have the same overall topology as the mtDNA present in the mouse mitochondrial network (i.e., circular monomers). Moreover, non-mtDNA replication and selection sequences required for maintaining the mitochondrial genomes in bacterial hosts are dispensable in these yeast mitochondria and could be efficiently and seamlessly removed by targeted homologous recombination within the mitochondria. These findings demonstrate that the yeast mtDNA replication system is capable of accurately replicating intact mammalian mtDNA genomes without sequence loss or rearrangement and that yeast mitochondria are a highly versatile host system for engineering complete mammalian mitochondrial genomes.

KW - Mitochondrial genome

KW - Mitochondrial host

KW - Non-mtDNA sequence

KW - Petite mutant

KW - Xenomitochondrial yeast

KW - mtDNA engineering

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SN - 1567-7249

VL - 46

SP - 15

EP - 21

JO - Mitochondrion

JF - Mitochondrion

ER -

Intramitochondrial transfer and engineering of mammalian mitochondrial genomes in yeast

Abstract

Bibliographical note

Keywords

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