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In Vitro-Reconstituted Nucleoids Can Block Mitochondrial DNA Replication and Transcription

Journal article
Authors G. Farge
Majda Mehmedovic
M. Baclayon
Smjl van den Wildenberg
W. H. Roos
Claes M Gustafsson
G. J. L. Wuite
Maria Falkenberg
Published in Cell Reports
Volume 8
Issue 1
Pages 66-74
ISSN 2211-1247
Publication year 2014
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 66-74
Language en
Links dx.doi.org/10.1016/j.celrep.2014.05...
Keywords COPY scigloo.NUMBER, FACTOR-A, FLUORESCENCE MICROSCOPY, MTDNA, TFAM, MAINTENANCE, INITIATION, PROMOTER, REVEALS, STRAND, Cell Biology
Subject categories Cell Biology

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, TFAMforms stable protein filaments onDNAthat 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.

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