Characterization of human cytomegalovirus-induced DNA polymerase and the associated 3'-to-5', exonuclease.
Characterization of human cytomegalovirus-induced DNA polymerase and the associated 3'-to-5', exonuclease.
复制标题
人巨细胞病毒诱导的 DNA 聚合酶和相关 3 至 5 核酸外切酶的表征。
DOI：
10.1016/0042-6822(83)90339-2
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发表时间：
1983
期刊：
影响因子：
3.7
通讯作者：
S. Yoshida
中科院分区：
文献类型：
--
作者：
Y. Nishiyama;K. Maeno;S. Yoshida
从宿主细胞DNA聚合酶中分离出人巨细胞病毒(HCMV)诱导的DNA聚合酶活性，并用磷酸纤维素柱层析和DNA-纤维素柱层析纯化。DNA聚合酶活性受膦乙酸、除草剂、araATP和N-乙基马来酰亚胺的强烈抑制，但对2‘，3’二脱氧TTP有抗性。巨细胞病毒诱导的DNA聚合酶对这些试剂的敏感性与宿主细胞DNA聚合酶α相似。然而，100mM硫酸铵可使人巨细胞病毒诱导的DNA聚合酶活性增加数倍，从而对α活性产生强烈的抑制作用。此外，还发现3‘-to DNA5’外切酶活性与−聚合酶密切相关。外切酶也被硫酸铵刺激，被磷酸醋酸抑制，并以单链DNA为底物。结果提示，3‘-to DNA5’外切酶可能作为−聚合酶的一个组成部分在聚合过程中起到校对作用。
A DNA polymerase activity induced by human cytomegalovirus (HCMV) was separated from host cell DNA polymerase and purified by phosphocellulose and DNA-cellulose column chromatography. The DNA polymerase activity was strongly inhibited by phosphonoacetic acid, aphidicolin, araATP, andN-ethylmaleimide, but it was resistant to 2′,3′dideoxyTTP. The sensitivity of HCMV-induced DNA polymerase to these reagents resembles that of host cell DNA polymerase α. However, HCMV-induced DNA polymerase activity was stimulated several fold by 100 mMammonium sulfate, by which DNA polymerase α activity was strongly inhibited. Furthermore, it was found that a 3′-to−5′ exonuclease activity was tightly associated with the HCMV-induced DNA polymerase. The exonuclease was also stimulated by ammonium sulfate, was inhibited by phosphoacetic acid, and it preferred single-stranded DNA as a substrate. The results suggest that the 3′-to−5′ exonuclease may play a role in proofreading in the polymerization process as an integral part of the HCMV-induced DNA polymerase.