Kaufmann G, Falk H H
Nucleic Acids Res. 1982 Apr 10;10(7):2309-21. doi: 10.1093/nar/10.7.2309.
A transient decaribonucleotide (iRNA) is covalently linked to nascent eukaryotic DNA chains at their 5' end. Searching for the putative iRNA polymerase (primase), we detected in extracts from SV40-infected cells a DNA-dependent incorporation of UMP residues from UTP into free and DNA linked deca- or similarly sized ribonucleotides. Denatured salmon sperm DNA served as the standard template in this reaction. SV40 FIII DNA was also an effective template, SV40 FII DNA was ineffective while FI yielded mainly free decaribonucleotides. The incorporation depended on the other rNTPs and was resistant to high concentrations of alpha-amanitin and rifamycin AF/013, drugs inhibitory to RNA polymerases I, II and III. The results implicate the decaribonucleotide polymerase in the priming of nascent DNA chains and suggest that the unique size of iRNA is encoded within its primase.
一种瞬时脱羧核苷酸(iRNA)在新生真核DNA链的5'末端与其共价连接。在寻找假定的iRNA聚合酶(引发酶)的过程中,我们在感染SV40的细胞提取物中检测到从UTP将UMP残基DNA依赖性掺入游离的以及与DNA相连的十聚或类似大小的核糖核苷酸中。变性的鲑鱼精DNA在该反应中用作标准模板。SV40 FIII DNA也是一种有效的模板,SV40 FII DNA无效,而FI主要产生游离的脱羧核苷酸。这种掺入依赖于其他rNTP,并且对高浓度的α-鹅膏蕈碱和利福霉素AF/013具有抗性,这两种药物对RNA聚合酶I、II和III具有抑制作用。结果表明脱羧核苷酸聚合酶参与新生DNA链的引发,并表明iRNA的独特大小是由其引发酶编码的。