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Mitochondrial RNA polymerase is needed for activation of the origin of light-strand DNA replication.

Journal article
Authors Javier Miralles Fusté
Sjoerd Wanrooij
Elisabeth Jemt
Caroline E Granycome
Tricia J Cluett
Yonghong Shi
Neli Atanassova
Ian J Holt
Claes M Gustafsson
Maria Falkenberg
Published in Molecular cell
Volume 37
Issue 1
Pages 67-78
ISSN 1097-4164
Publication year 2010
Published at Institute of Biomedicine
Pages 67-78
Language en
Keywords DNA Replication, DNA, Mitochondrial, biosynthesis, chemistry, DNA-Directed RNA Polymerases, physiology, Gene Silencing, Humans, Models, Genetic, Nucleic Acid Conformation, Poly T, chemistry, Replication Origin
Subject categories Chemistry


Mitochondrial DNA is replicated by a unique enzymatic machinery, which is distinct from the replication apparatus used for copying the nuclear genome. We examine here the mechanisms of origin-specific initiation of lagging-strand DNA synthesis in human mitochondria. We demonstrate that the mitochondrial RNA polymerase (POLRMT) is the primase required for initiation of DNA synthesis from the light-strand origin of DNA replication (OriL). Using only purified POLRMT and DNA replication factors, we can faithfully reconstitute OriL-dependent initiation in vitro. Leading-strand DNA synthesis is initiated from the heavy-strand origin of DNA replication and passes OriL. The single-stranded OriL is exposed and adopts a stem-loop structure. At this stage, POLRMT initiates primer synthesis from a poly-dT stretch in the single-stranded loop region. After about 25 nt, POLRMT is replaced by DNA polymerase gamma, and DNA synthesis commences. Our findings demonstrate that POLRMT can function as an origin-specific primase in mammalian mitochondria.

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