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J. Biol. Chem., Vol. 260, Issue 21, 11530-11535, Sep, 1985
TW Wong and DA Clayton
A family of enzymatic activities isolated from human mitochondria is
capable of initiating DNA replication on single-stranded templates. The
principal enzymes include at least a primase and DNA polymerase gamma and
require that rNTPs as well as dNTPs be present in the reaction mixture.
Poly(dC) and poly(dT), as well as M13 phage DNA, are excellent templates
for the primase activity. A single-stranded DNA containing the cloned
origin of mitochondrial light-strand synthesis can be a more efficient
template than M13 phage DNA alone. Primase and DNA polymerase activities
were separated from each other by sedimentation in a glycerol density
gradient. Using M13 phage DNA as template, these mitochondrial enzymes
synthesize RNA primers that are 9 to 12 nucleotides in size and are
covalently linked to nascent DNA. The formation of primers appears to be
the rate-limiting step in the replication process. Replication of M13 DNA
is sensitive to N- ethylmaleimide and dideoxynucleoside triphosphates, but
insensitive to rifampicin, alpha-amanitin, and aphidicolin.
Isolation and characterization of a DNA primase from human mitochondria
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