In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription

Géraldine Farge; Majda Mehmedovic; Marian Baclayon; Siet M J L van den Wildenberg; Wouter H Roos; Claes M Gustafsson; Gijs J L Wuite; Maria Falkenberg

doi:10.1016/j.celrep.2014.05.046

In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription

Géraldine Farge, Majda Mehmedovic, Marian Baclayon, Siet M J L van den Wildenberg, Wouter H Roos, Claes M Gustafsson, Gijs J L Wuite, Maria Falkenberg

Molecular Biophysics

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

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Abstract

The mechanisms regulating the number of active copies of mtDNA are still unclear. A mammalian cell typically contains 1,000-10,000 copies of mtDNA, which are packaged into nucleoprotein complexes termed nucleoids. The main protein component of these structures is mitochondrial transcription factor A (TFAM). Here, we reconstitute nucleoid-like particles in vitro and demonstrate that small changes in TFAM levels dramatically impact the fraction of DNA molecules available for transcription and DNA replication. Compaction by TFAM is highly cooperative, and at physiological ratios of TFAM to DNA, there are large variations in compaction, from fully compacted nucleoids to naked DNA. In compacted nucleoids, TFAM forms stable protein filaments on DNA that block melting and prevent progression of the replication and transcription machineries. Based on our observations, we suggest that small variations in the TFAM-to-mtDNA ratio may be used to regulate mitochondrial gene transcription and DNA replication.

Original language	English
Pages (from-to)	66-74
Number of pages	9
Journal	Cell reports
Volume	8
Issue number	1
DOIs	https://doi.org/10.1016/j.celrep.2014.05.046
Publication status	Published - 10-Jul-2014

Keywords

DNA Replication
DNA, Mitochondrial
DNA, Viral
Humans
Protein Binding
Protein Multimerization
Transcription Factors
Transcription, Genetic
COPY NUMBER
FACTOR-A
FLUORESCENCE MICROSCOPY
MTDNA
TFAM
MAINTENANCE
INITIATION
PROMOTER
REVEALS
STRAND

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In Vitro-Reconstituted Nucleoids Can Block Mitochondrial DNA Replication and TranscriptionFinal publisher's version, 2.21 MBLicence: CC BY

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title = "In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription",

abstract = "The mechanisms regulating the number of active copies of mtDNA are still unclear. A mammalian cell typically contains 1,000-10,000 copies of mtDNA, which are packaged into nucleoprotein complexes termed nucleoids. The main protein component of these structures is mitochondrial transcription factor A (TFAM). Here, we reconstitute nucleoid-like particles in vitro and demonstrate that small changes in TFAM levels dramatically impact the fraction of DNA molecules available for transcription and DNA replication. Compaction by TFAM is highly cooperative, and at physiological ratios of TFAM to DNA, there are large variations in compaction, from fully compacted nucleoids to naked DNA. In compacted nucleoids, TFAM forms stable protein filaments on DNA that block melting and prevent progression of the replication and transcription machineries. Based on our observations, we suggest that small variations in the TFAM-to-mtDNA ratio may be used to regulate mitochondrial gene transcription and DNA replication.",

keywords = "DNA Replication, DNA, Mitochondrial, DNA, Viral, Humans, Protein Binding, Protein Multimerization, Transcription Factors, Transcription, Genetic, COPY NUMBER, FACTOR-A, FLUORESCENCE MICROSCOPY, MTDNA, TFAM, MAINTENANCE, INITIATION, PROMOTER, REVEALS, STRAND",

author = "G{\'e}raldine Farge and Majda Mehmedovic and Marian Baclayon and {van den Wildenberg}, {Siet M J L} and Roos, {Wouter H} and Gustafsson, {Claes M} and Wuite, {Gijs J L} and Maria Falkenberg",

year = "2014",

month = jul,

day = "10",

doi = "10.1016/j.celrep.2014.05.046",

language = "English",

volume = "8",

pages = "66--74",

journal = "Cell reports",

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TY - JOUR

T1 - In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription

AU - Farge, Géraldine

AU - Mehmedovic, Majda

AU - Baclayon, Marian

AU - van den Wildenberg, Siet M J L

AU - Roos, Wouter H

AU - Gustafsson, Claes M

AU - Wuite, Gijs J L

AU - Falkenberg, Maria

PY - 2014/7/10

Y1 - 2014/7/10

N2 - The mechanisms regulating the number of active copies of mtDNA are still unclear. A mammalian cell typically contains 1,000-10,000 copies of mtDNA, which are packaged into nucleoprotein complexes termed nucleoids. The main protein component of these structures is mitochondrial transcription factor A (TFAM). Here, we reconstitute nucleoid-like particles in vitro and demonstrate that small changes in TFAM levels dramatically impact the fraction of DNA molecules available for transcription and DNA replication. Compaction by TFAM is highly cooperative, and at physiological ratios of TFAM to DNA, there are large variations in compaction, from fully compacted nucleoids to naked DNA. In compacted nucleoids, TFAM forms stable protein filaments on DNA that block melting and prevent progression of the replication and transcription machineries. Based on our observations, we suggest that small variations in the TFAM-to-mtDNA ratio may be used to regulate mitochondrial gene transcription and DNA replication.

AB - The mechanisms regulating the number of active copies of mtDNA are still unclear. A mammalian cell typically contains 1,000-10,000 copies of mtDNA, which are packaged into nucleoprotein complexes termed nucleoids. The main protein component of these structures is mitochondrial transcription factor A (TFAM). Here, we reconstitute nucleoid-like particles in vitro and demonstrate that small changes in TFAM levels dramatically impact the fraction of DNA molecules available for transcription and DNA replication. Compaction by TFAM is highly cooperative, and at physiological ratios of TFAM to DNA, there are large variations in compaction, from fully compacted nucleoids to naked DNA. In compacted nucleoids, TFAM forms stable protein filaments on DNA that block melting and prevent progression of the replication and transcription machineries. Based on our observations, we suggest that small variations in the TFAM-to-mtDNA ratio may be used to regulate mitochondrial gene transcription and DNA replication.

KW - DNA Replication

KW - DNA, Mitochondrial

KW - DNA, Viral

KW - Humans

KW - Protein Binding

KW - Protein Multimerization

KW - Transcription Factors

KW - Transcription, Genetic

KW - COPY NUMBER

KW - FACTOR-A

KW - FLUORESCENCE MICROSCOPY

KW - MTDNA

KW - TFAM

KW - MAINTENANCE

KW - INITIATION

KW - PROMOTER

KW - REVEALS

KW - STRAND

U2 - 10.1016/j.celrep.2014.05.046

DO - 10.1016/j.celrep.2014.05.046

M3 - Article

C2 - 24981867

SN - 2211-1247

VL - 8

SP - 66

EP - 74

JO - Cell reports

JF - Cell reports

IS - 1

ER -

In vitro-reconstituted nucleoids can block mitochondrial DNA replication and transcription

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Keywords

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