McMillan S, Edenberg H J, Radany E H, Friedberg R C, Friedberg E C
J Virol. 1981 Oct;40(1):211-23. doi: 10.1128/JVI.40.1.211-223.1981.
Recent studies have shown purified preparations of phage T4 UV DNA-incising activity (T4 UV endonuclease or endonuclease V of phage T4) contain a pyrimidine dimer-DNA glycosylase activity that catalyzes hydrolysis of the 5' glycosyl bond of dimerized pyrimidines in UV-irradiated DNA. Such enzyme preparations have also been shown to catalyze the hydrolysis of phosphodiester bonds in UV-irradiated DNA at a neutral pH, presumably reflecting the action of an apurinic/apyrimidinic endonuclease at the apyrimidinic sites created by the pyrimidine dimer-DNA glycosylase. In this study we found that preparations of T4 UV DNA-incising activity contained apurinic/apyrimidinic endonuclease activity that nicked depurinated form I simian virus 40 DNA. Apurinic/apyrimidinic endonuclease activity was also found in extracts of Escherichia coli infected with T4 denV+ phage. Extracts of cells infected with T4 denV mutants contained significantly lower levels of apurinic/apyrimidinic endonuclease activity; these levels were no greater than the levels present in extracts of uninfected cells. Furthermore, the addition of DNA containing apurinic or apyrimidinic sites to reactions containing UV-irradiated DNA and T4 enzyme resulted in competition for pyrimidine dimer-DNA glycosylase activity against the UV-irradiated DNA. On the basis of these results, we concluded that apurinic/apyrimidinic endonuclease activity is encoded by the denV gene of phage T4, the same gene that codes for pyrimidine dimer-DNA glycosylase activity.
最近的研究表明,纯化的噬菌体T4紫外线切割DNA活性制剂(T4紫外线内切酶或噬菌体T4的内切酶V)含有嘧啶二聚体-DNA糖基化酶活性,该活性催化紫外线照射的DNA中嘧啶二聚体的5'糖苷键水解。此类酶制剂还被证明在中性pH下能催化紫外线照射的DNA中磷酸二酯键的水解,这大概反映了脱嘌呤/脱嘧啶内切酶在由嘧啶二聚体-DNA糖基化酶产生的脱嘧啶位点的作用。在本研究中,我们发现T4紫外线切割DNA活性制剂含有脱嘌呤/脱嘧啶内切酶活性,该活性能切割去嘌呤的I型猿猴病毒40 DNA。在感染了T4 denV⁺噬菌体的大肠杆菌提取物中也发现了脱嘌呤/脱嘧啶内切酶活性。感染T4 denV突变体的细胞提取物中脱嘌呤/脱嘧啶内切酶活性水平显著降低;这些水平不高于未感染细胞提取物中的水平。此外,向含有紫外线照射的DNA和T4酶的反应中添加含有脱嘌呤或脱嘧啶位点的DNA,会导致嘧啶二聚体-DNA糖基化酶活性与紫外线照射的DNA竞争。基于这些结果,我们得出结论,脱嘌呤/脱嘧啶内切酶活性由噬菌体T4的denV基因编码,该基因与编码嘧啶二聚体-DNA糖基化酶活性的基因相同。