含尿嘧啶DNA的酶促降解。II. 枯草芽孢杆菌未分级提取物中N-糖苷酶和核酸酶活性的证据。
Enzymatic degradation of uracil-containing DNA. II. Evidence for N-glycosidase and nuclease activities in unfractionated extracts of Bacillus subtilis.
作者信息
Duncan J, Hamilton L, Friedberg E C
出版信息
J Virol. 1976 Aug;19(2):338-45. doi: 10.1128/JVI.19.2.338-345.1976.
Abstract
Further studies have confirmed our earlier observations that in the presence of EDTA, degradation of phage PBS2 [3H]uracil-labeled DNA is effected by an N-glycosidase activity in extracts of Bacillus subtilis that removes free uracil from DNA. In addition, such extracts contain a nuclease activity that attacks PBS2 DNA in the presence of CaCl2. The nuclease activity is not observed under conditions that inactivate N-glycosidase activity but does attack DNA that has been preincubated to remove uracil by N-glycosidase action. We therefore postulate that the nuclease requires N-glycosidase action to generate substrate for its activity, i.e., the nuclease appears to attack depyrimidinated sites rather than uracil sites in phage PBS2 DNA.
摘要
进一步的研究证实了我们早期的观察结果，即在乙二胺四乙酸（EDTA）存在的情况下，枯草芽孢杆菌提取物中的N-糖苷酶活性会导致噬菌体PBS2 [3H]尿嘧啶标记的DNA降解，该活性会从DNA中去除游离尿嘧啶。此外，此类提取物含有一种核酸酶活性，在氯化钙存在的情况下会攻击PBS2 DNA。在使N-糖苷酶活性失活的条件下未观察到核酸酶活性，但它确实会攻击经预孵育以通过N-糖苷酶作用去除尿嘧啶的DNA。因此，我们推测核酸酶需要N-糖苷酶作用来生成其活性的底物，即核酸酶似乎攻击噬菌体PBS2 DNA中的脱嘧啶位点而非尿嘧啶位点。
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