Savinkova L K, Sokolenko A A, Tulokhonov I I, Knoppe V L, Salganik R I, Ven'iaminova A G, Repkova M N, Komarova N I
Mol Biol (Mosk). 1993 Jan-Feb;27(1):64-71.
It was shown previously that E. coli RNA polymerase in a highly selective manner recognizes and binds 11-14-mere oligodeoxyribonucleotides related to the "-10" region of the nontranscribed DNA strand of bacterial gene promoters. The oligodeoxyribonucleotides cover the Pribnow box with flanking nucleotides up to the transcription start. These affinity oligodeoxyribonucleotides inhibit competitively the transcription of bacterial DNA carried out by E. coli RNA polymerase. The present work has demonstrated that E. coli RNA polymerase is not capable of binding the oligoribonucleotides homologous to the affinity oligodeoxyribonucleotides related to the "-10" area of the spc promoter, but binds the oligoribonucleotides which are complementary to the latter. The oligoribonucleotides with a high affinity for the E. coli RNA polymerase strongly inhibit transcription of the bacterial DNA. Attachment of alkylating groups to the 5'-ends of the affinity oligodeoxy- and oligoribonucleotides provides their covalent binding to the E. coli RNA polymerase subunits. It was shown that the modified affinity 32P-labelled oligodeoxyribonucleotide is covalently bound to the sigma-subunit while the modified affinity 32P-labelled oligoribonucleotide is covalently bound to the beta'beta-subunits of the E. coli RNA polymerase. It is suggested that the affinity oligoribonucleotides can be transcribed from the non-transcribed DNA strand in the region of the open complex and functions presumably as a primer which is splitted later from the nascent RNA or as a regulator of transcription.
先前的研究表明,大肠杆菌RNA聚合酶以高度选择性的方式识别并结合与细菌基因启动子非转录DNA链“-10”区域相关的11至14聚体寡脱氧核糖核苷酸。这些寡脱氧核糖核苷酸覆盖普里布诺框及其侧翼核苷酸直至转录起始点。这些亲和性寡脱氧核糖核苷酸竞争性抑制大肠杆菌RNA聚合酶对细菌DNA的转录。目前的研究表明,大肠杆菌RNA聚合酶不能结合与spc启动子“-10”区域相关的亲和性寡脱氧核糖核苷酸同源的寡核糖核苷酸,但能结合与后者互补的寡核糖核苷酸。对大肠杆菌RNA聚合酶具有高亲和力的寡核糖核苷酸强烈抑制细菌DNA的转录。将烷基化基团连接到亲和性寡脱氧核糖核苷酸和寡核糖核苷酸的5'端可使其与大肠杆菌RNA聚合酶亚基共价结合。结果表明,修饰后的亲和性32P标记寡脱氧核糖核苷酸与σ亚基共价结合,而修饰后的亲和性32P标记寡核糖核苷酸与大肠杆菌RNA聚合酶的β'β亚基共价结合。有人提出,亲和性寡核糖核苷酸可在开放复合物区域从非转录DNA链转录而来,其功能可能是作为引物,随后从新生RNA中分离,或作为转录调节因子。