Nuclear and mitochondrial DNA-dependent RNA polymerases in bovine spermatozoa

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Abstract
DNA-dependent RNA polymerases were solubilized from separated head and tail fractions from normal bovine spermatozoa and from spermatozoa carrying the decapitated sperm defect. When enzyme extracts from separated heads and tails were chromatographed on DEAE-Sephadex, the head fraction was resolved into 2 distinguishable peaks eluting at .apprx. 0.11 and 0.15 M (NH4)2SO4 while the tail fraction yielded 4 distinct peaks eluting at about 0.11, 0.15, 0.255 and 0.35 M (NH4)2SO4. Results identical to those observed for sperm tails were obtained with extracts prepared from highly purified mitochondria from bovine or murine heart or liver. Optimization of reaction parameters and inhibitor studies with .alpha.-amanitin and rifampicin revealed strong similarities between eucaryotic nuclear RNA polymerases 1 and 2 and the 2 RNA polymerases associated with sperm heads. Similar experiments comparing the RNA polymerases from somatic mitochondria and sperm tails suggested the sperm tail enzymes were mitochondrial in origin.