Hecht N B, Williams J L
J Reprod Fertil. 1979 Sep;57(1):157-65. doi: 10.1530/jrf.0.0570157.
DNA-dependent RNA polymerases have been 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 about 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 indentical 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.
已从正常牛精子以及携带“断头精子缺陷”的精子的分离头部和尾部组分中溶解出依赖DNA的RNA聚合酶。当将分离的头部和尾部的酶提取物在DEAE-葡聚糖凝胶上进行色谱分析时,头部组分被分离为两个可区分的峰,分别在约0.11和0.15M硫酸铵处洗脱,而尾部组分产生4个不同的峰,分别在约0.11、0.15、0.255和0.35M硫酸铵处洗脱。用从牛或鼠心脏或肝脏的高度纯化线粒体制备的提取物获得了与精子尾部观察到的结果相同的结果。用α-鹅膏蕈碱和利福平对反应参数进行优化以及抑制剂研究表明,真核细胞核RNA聚合酶1和2与与精子头部相关的2种RNA聚合酶之间存在很强的相似性。比较体细胞线粒体和精子尾部的RNA聚合酶的类似实验表明,精子尾部的酶起源于线粒体。