The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells

M. Di Re, H. Sembongi, J. He, A. Reyes, Takehiro Yasukawa, P. Martinsson, L. J. Bailey, S. Goffart, J. D. Boyd-kirkup, T. S. Wong, A. R. Fersht, J. N. Spelbrink, I. J. Holt

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The accessory subunit of mitochondrial DNA polymerase γ, POLGβ, functions as a processivity factor in vitro. Here we show POLGβ has additional roles in mitochondrial DNA metabolism. Mitochondrial DNA is arranged in nucleoprotein complexes, or nucleoids, which often contain multiple copies of the mitochondrial genome. Gene-silencing of POLGβ increased nucleoid numbers, whereas overexpression of POLGβ reduced the number and increased the size of mitochondrial nucleoids. Both increased and decreased expression of POLGβ altered nucleoid structure and precipitated a marked decrease in 7S DNA molecules, which form short displacement-loops on mitochondrial DNA. Recombinant POLGβ preferentially bound to plasmids with a short displacement-loop, in contrast to POLGα. These findings support the view that the mitochondrial D-loop acts as a protein recruitment centre, and suggest POLGβ is a key factor in the organization of mitochondrial DNA in multigenomic nucleoprotein complexes.

Original languageEnglish
Article numbergkp614
Pages (from-to)5701-5713
Number of pages13
JournalNucleic acids research
Volume37
Issue number17
DOIs
Publication statusPublished - Jul 22 2009
Externally publishedYes

Fingerprint

DNA-Directed DNA Polymerase
Mitochondrial DNA
Cultured Cells
Nucleoproteins
Mitochondrial Size
Mitochondrial Genome
Gene Silencing
Plasmids
DNA
Proteins

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells. / Di Re, M.; Sembongi, H.; He, J.; Reyes, A.; Yasukawa, Takehiro; Martinsson, P.; Bailey, L. J.; Goffart, S.; Boyd-kirkup, J. D.; Wong, T. S.; Fersht, A. R.; Spelbrink, J. N.; Holt, I. J.

In: Nucleic acids research, Vol. 37, No. 17, gkp614, 22.07.2009, p. 5701-5713.

Research output: Contribution to journalArticle

Di Re, M, Sembongi, H, He, J, Reyes, A, Yasukawa, T, Martinsson, P, Bailey, LJ, Goffart, S, Boyd-kirkup, JD, Wong, TS, Fersht, AR, Spelbrink, JN & Holt, IJ 2009, 'The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells', Nucleic acids research, vol. 37, no. 17, gkp614, pp. 5701-5713. https://doi.org/10.1093/nar/gkp614
Di Re, M. ; Sembongi, H. ; He, J. ; Reyes, A. ; Yasukawa, Takehiro ; Martinsson, P. ; Bailey, L. J. ; Goffart, S. ; Boyd-kirkup, J. D. ; Wong, T. S. ; Fersht, A. R. ; Spelbrink, J. N. ; Holt, I. J. / The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells. In: Nucleic acids research. 2009 ; Vol. 37, No. 17. pp. 5701-5713.
@article{40dde85a73854fe2bc61b82db9c1566c,
title = "The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells",
abstract = "The accessory subunit of mitochondrial DNA polymerase γ, POLGβ, functions as a processivity factor in vitro. Here we show POLGβ has additional roles in mitochondrial DNA metabolism. Mitochondrial DNA is arranged in nucleoprotein complexes, or nucleoids, which often contain multiple copies of the mitochondrial genome. Gene-silencing of POLGβ increased nucleoid numbers, whereas overexpression of POLGβ reduced the number and increased the size of mitochondrial nucleoids. Both increased and decreased expression of POLGβ altered nucleoid structure and precipitated a marked decrease in 7S DNA molecules, which form short displacement-loops on mitochondrial DNA. Recombinant POLGβ preferentially bound to plasmids with a short displacement-loop, in contrast to POLGα. These findings support the view that the mitochondrial D-loop acts as a protein recruitment centre, and suggest POLGβ is a key factor in the organization of mitochondrial DNA in multigenomic nucleoprotein complexes.",
author = "{Di Re}, M. and H. Sembongi and J. He and A. Reyes and Takehiro Yasukawa and P. Martinsson and Bailey, {L. J.} and S. Goffart and Boyd-kirkup, {J. D.} and Wong, {T. S.} and Fersht, {A. R.} and Spelbrink, {J. N.} and Holt, {I. J.}",
year = "2009",
month = "7",
day = "22",
doi = "10.1093/nar/gkp614",
language = "English",
volume = "37",
pages = "5701--5713",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "17",

}

TY - JOUR

T1 - The accessory subunit of mitochondrial DNA polymerase γ determines the DNA content of mitochondrial nucleoids in human cultured cells

AU - Di Re, M.

AU - Sembongi, H.

AU - He, J.

AU - Reyes, A.

AU - Yasukawa, Takehiro

AU - Martinsson, P.

AU - Bailey, L. J.

AU - Goffart, S.

AU - Boyd-kirkup, J. D.

AU - Wong, T. S.

AU - Fersht, A. R.

AU - Spelbrink, J. N.

AU - Holt, I. J.

PY - 2009/7/22

Y1 - 2009/7/22

N2 - The accessory subunit of mitochondrial DNA polymerase γ, POLGβ, functions as a processivity factor in vitro. Here we show POLGβ has additional roles in mitochondrial DNA metabolism. Mitochondrial DNA is arranged in nucleoprotein complexes, or nucleoids, which often contain multiple copies of the mitochondrial genome. Gene-silencing of POLGβ increased nucleoid numbers, whereas overexpression of POLGβ reduced the number and increased the size of mitochondrial nucleoids. Both increased and decreased expression of POLGβ altered nucleoid structure and precipitated a marked decrease in 7S DNA molecules, which form short displacement-loops on mitochondrial DNA. Recombinant POLGβ preferentially bound to plasmids with a short displacement-loop, in contrast to POLGα. These findings support the view that the mitochondrial D-loop acts as a protein recruitment centre, and suggest POLGβ is a key factor in the organization of mitochondrial DNA in multigenomic nucleoprotein complexes.

AB - The accessory subunit of mitochondrial DNA polymerase γ, POLGβ, functions as a processivity factor in vitro. Here we show POLGβ has additional roles in mitochondrial DNA metabolism. Mitochondrial DNA is arranged in nucleoprotein complexes, or nucleoids, which often contain multiple copies of the mitochondrial genome. Gene-silencing of POLGβ increased nucleoid numbers, whereas overexpression of POLGβ reduced the number and increased the size of mitochondrial nucleoids. Both increased and decreased expression of POLGβ altered nucleoid structure and precipitated a marked decrease in 7S DNA molecules, which form short displacement-loops on mitochondrial DNA. Recombinant POLGβ preferentially bound to plasmids with a short displacement-loop, in contrast to POLGα. These findings support the view that the mitochondrial D-loop acts as a protein recruitment centre, and suggest POLGβ is a key factor in the organization of mitochondrial DNA in multigenomic nucleoprotein complexes.

UR - http://www.scopus.com/inward/record.url?scp=73049113711&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=73049113711&partnerID=8YFLogxK

U2 - 10.1093/nar/gkp614

DO - 10.1093/nar/gkp614

M3 - Article

VL - 37

SP - 5701

EP - 5713

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 17

M1 - gkp614

ER -