ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEIC ACID .3. THE INCORPORATION OF PYRIMIDINE AND PURINE ANALOGUES INTO DEOXYRIBONUCLEIC ACID
ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEIC ACID .3. THE INCORPORATION OF PYRIMIDINE AND PURINE ANALOGUES INTO DEOXYRIBONUCLEIC ACID
复制标题
DOI：
10.1073/pnas.44.7.633
复制
发表时间：
1958-01-01
影响因子：
11.1
通讯作者：
KORNBERG, A
中科院分区：
文献类型：
--
作者：
BESSMAN, MJ;LEHMAN, IR;KORNBERG, A
嘧啶和嘌呤碱基的类似物的脱氧核苷三磷酸被化学地制备，并且被测试为来自大肠杆菌的酶的底物，该酶将脱氧核糖核苷酸聚合成脱氧核糖核酸（DNA）。尿嘧啶和5-溴尿嘧啶取代胸腺嘧啶，5-甲基和5-溴胞嘧啶取代胞嘧啶，次黄嘌呤取代鸟嘌呤。黄嘌呤没有掺入DNA中。对含尿嘧啶产物的分析表明，尿嘧啶以3[image]-5[image]磷酸二酯键与腺嘌呤、鸟嘌呤、胞嘧啶和尿嘧啶的脱氧核苷酸结合。这些类似物对天然碱基的特异性替代，为沃森和克里克提出的DNA结构双螺旋中的碱基配对关系提供了额外的支持。E.将5-溴脱氧尿苷酸β-磷酸化为三磷酸的大肠杆菌中的磷酸化酶和脱氧尿苷酸磷酸化的这种激酶的明显缺乏与细胞将5-溴尿嘧啶而不是尿嘧啶掺入DNA中的事实一致。
The deoxyribonucleoside triphosphates of analogs of the pyrimidine and purine bases were prepared chemically and tested as substrates for the enzyme from Escherichia coli which polymerizes deoxyribonucleotides to deoxyribonucleic acid (DNA). Uracil and 5-bromouracil were incorporated into DNA specifically in place of thymine; 5-methyl-and 5-bromocytosine in place of cytosine; and hypoxanthine in place of guanine. Xanthine was not incorporated into DNA. Analysis of the uracil-containing product demonstrated that the uracil was bound in 3[image]-5[image] phosphodiester linkage with each of the deoxyribotides of adenine, guanine, cytosine, and uracil. The specific replacement of the natural bases by these analogs offers additional support for the base-pairing relationships in the double helix proposed by Watson and Crick for the structure of DNA. The existence of kinases in E. coli which phos-phorylate 5-bromodeoxyuridylate to the triphosphate and the apparent absence of such kinases for the phosphorylation of deoxyuridylate are in keeping with the fact that cells incorporate 5-bromouracil, but not uracil, into DNA.