Lee S K, Fuchs M S
Department of Biological Sciences, University of Notre Dame, IN 46556.
FEBS Lett. 1993 Feb 1;316(3):261-3. doi: 10.1016/0014-5793(93)81304-i.
The mosquito homolog of mammalian DNA polymerase epsilon, formerly known as a proliferating cell nuclear antigen (PCNA)-independent form of DNA polymerase delta, has been purified from mosquito larval extracts. The polymerase epsilon was separated from DNA polymerase alpha by chromatography on hydroxylapatite, and the enzyme was subsequently purified on single-stranded DNA agarose, followed by a 5' AMP-agarose chromatography step. The purified polymerase exhibits an intrinsic 3'-5' exonuclease activity and shows high activity using an oligo-primed DNA template. Neither human nor Drosophila PCNA stimulated this polymerase activity. Additional immunochemical and biochemical evidence indicates that this enzyme is distinct from DNA polymerase alpha.
哺乳动物DNA聚合酶ε的蚊子同源物,以前被称为增殖细胞核抗原(PCNA)非依赖性形式的DNA聚合酶δ,已从蚊子幼虫提取物中纯化出来。通过羟基磷灰石色谱法将聚合酶ε与DNA聚合酶α分离,随后该酶在单链DNA琼脂糖上纯化,接着进行5' AMP-琼脂糖色谱步骤。纯化后的聚合酶具有内在的3'-5'核酸外切酶活性,并且使用寡核苷酸引发的DNA模板时显示出高活性。人源和果蝇的PCNA均未刺激这种聚合酶活性。更多的免疫化学和生化证据表明,这种酶与DNA聚合酶α不同。