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Mammalian Mitochondrial DNA Replication Intermediates Are Essentially Duplex but Contain Extensive Tracts of RNA/DNA Hybrid |
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| Authors: | Jaakko LO Pohjoismäki J Bradley Holmes Stuart R Wood Takehiro Yasukawa Aurelio Reyes Tricia J Cluett Smaranda Willcox Andrew P Jackson Jack D Griffith Howard T Jacobs |
| Institution: | 1 Department of Forensic Medicine and Tampere University Hospital, FI-33014 University of Tampere, Finland 2 Institute of Medical Technology and Tampere University Hospital, FI-33014 University of Tampere, Finland 3 MRC-Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge CB2 0XY, UK 4 Program in Genomics of Differentiation, Eunice Kennedy Shriver National Institute of Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA 5 Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Mason Farm Road, Chapel Hill, NC 27599, USA 6 MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK |
| Abstract: | We demonstrate, using transmission electron microscopy and immunopurification with an antibody specific for RNA/DNA hybrid, that intact mitochondrial DNA replication intermediates are essentially duplex throughout their length but contain extensive RNA tracts on one strand. However, the extent of preservation of RNA in such molecules is highly dependent on the preparative method used. These findings strongly support the strand-coupled model of mitochondrial DNA replication involving RNA incorporation throughout the lagging strand. |
| Keywords: | mtRI mitochondrial replication intermediate RITOLS RNA incorporation throughout the lagging strand mtDNA mitochondrial DNA ssDNA single-stranded DNA TEM transmission electron microscopy 2D-AGE two-dimensional agarose gel electrophoresis AFM atomic force microscopy SSB single-stranded binding protein SDM strand-displacement model CsCl cesium chloride EMSA electrophoretic mobility shift assay |
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