2.50
Hdl Handle:
http://hdl.handle.net/10547/228721
Title:
Mitochondrial DNA replication during differentiation of murine embryonic stem cells
Authors:
Facucho-Oliveira, J.M.; Alderson, Jon; Spikings, Emma ( 0000-0001-8387-2098 ) ; Egginton, S.; St John, Justin C.
Abstract:
Oxidative phosphorylation (OXPHOS), the intracellular process that generates the majority of the ATP of a cell through the electron-transfer chain, is highly dependent on proteins encoded by the mitochondrial genome (mtDNA). MtDNA replication is regulated by the nuclear-encoded mitochondrial transcription factor A (TFAM) and the mitochondrial-specific DNA polymerase gamma, which consists of a catalytic (POLG) and an accessory (POLG2)subunit. Differentiation of pluripotent embryonic stem cells (ESCs) into specific cell types requires expansion of discrete populations of mitochondria and mtDNA replication to meet the specific metabolic requirements of the cell. We determined by real-time PCR that expression of pluripotent markers is reduced before the upregulation of Polg, Polg2 and Tfam in spontaneously differentiating R1 murine (m)ESCs, along with transient increases in mtDNA copy number. In D3 mESCs, the initial transient increase did not take place. However, precursors of neuronal and cardiomyocyte differentiation were positive for both POLG and TFAM. Similar-stage ESCs also showed active mtDNA replication, identified by 5-bromo-2 -deoxy-uridine labelling, as mtDNA copy number increased. Retinoic-acidinduced differentiation resulted in more consistent patterns of replication and upregulation of Polg, Polg2 and Tfam, whereas siRNA knockdown demonstrated that steadystate expression of POLG is essential for maintaining pluripotency.
Affiliation:
University of Birmingham
Citation:
Facucho-Oliveira, J.M., Alderson, J., Spikings, E., Egginton, S., St John, J.C. (2007) 'Mitochondrial DNA replication during differentiation of murine embryonic stem cells' Journal of Cell Science 120 (22):4025-4034
Publisher:
Company of Biologists
Journal:
Journal of Cell Science
Issue Date:
15-Nov-2007
URI:
http://hdl.handle.net/10547/228721
DOI:
10.1242/jcs.016972
Additional Links:
http://jcs.biologists.org/cgi/doi/10.1242/jcs.016972
Type:
Article
Language:
en
ISSN:
0021-9533; 1477-9137
Appears in Collections:
Cell and Cryobiology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorFacucho-Oliveira, J.M.en_GB
dc.contributor.authorAlderson, Jonen_GB
dc.contributor.authorSpikings, Emmaen_GB
dc.contributor.authorEgginton, S.en_GB
dc.contributor.authorSt John, Justin C.en_GB
dc.date.accessioned2012-06-13T10:18:39Zen
dc.date.available2012-06-13T10:18:39Zen
dc.date.issued2007-11-15en
dc.identifier.citationFacucho-Oliveira, J.M., Alderson, J., Spikings, E., Egginton, S., St John, J.C. (2007) 'Mitochondrial DNA replication during differentiation of murine embryonic stem cells' Journal of Cell Science 120 (22):4025-4034en_GB
dc.identifier.issn0021-9533en
dc.identifier.issn1477-9137en
dc.identifier.doi10.1242/jcs.016972en
dc.identifier.urihttp://hdl.handle.net/10547/228721en
dc.description.abstractOxidative phosphorylation (OXPHOS), the intracellular process that generates the majority of the ATP of a cell through the electron-transfer chain, is highly dependent on proteins encoded by the mitochondrial genome (mtDNA). MtDNA replication is regulated by the nuclear-encoded mitochondrial transcription factor A (TFAM) and the mitochondrial-specific DNA polymerase gamma, which consists of a catalytic (POLG) and an accessory (POLG2)subunit. Differentiation of pluripotent embryonic stem cells (ESCs) into specific cell types requires expansion of discrete populations of mitochondria and mtDNA replication to meet the specific metabolic requirements of the cell. We determined by real-time PCR that expression of pluripotent markers is reduced before the upregulation of Polg, Polg2 and Tfam in spontaneously differentiating R1 murine (m)ESCs, along with transient increases in mtDNA copy number. In D3 mESCs, the initial transient increase did not take place. However, precursors of neuronal and cardiomyocyte differentiation were positive for both POLG and TFAM. Similar-stage ESCs also showed active mtDNA replication, identified by 5-bromo-2 -deoxy-uridine labelling, as mtDNA copy number increased. Retinoic-acidinduced differentiation resulted in more consistent patterns of replication and upregulation of Polg, Polg2 and Tfam, whereas siRNA knockdown demonstrated that steadystate expression of POLG is essential for maintaining pluripotency.en_GB
dc.language.isoenen
dc.publisherCompany of Biologistsen_GB
dc.relation.urlhttp://jcs.biologists.org/cgi/doi/10.1242/jcs.016972en_GB
dc.rightsArchived with thanks to Journal of Cell Scienceen_GB
dc.subjectmitochondrial DNAen_GB
dc.subjectreplicationen
dc.subjectPOLGen
dc.subjectTFAMen
dc.subjectembryonic stem cellsen
dc.titleMitochondrial DNA replication during differentiation of murine embryonic stem cellsen
dc.typeArticleen
dc.contributor.departmentUniversity of Birminghamen_GB
dc.identifier.journalJournal of Cell Scienceen_GB
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