Slippage at the initiation of RNA synthesis by Qβ replicase results in a periodic polyG pattern

Авторы: Lobodin Kirill V., Chetverina Helena V., Chetverin Alexander B.
Журнал: FEBS Letters
Год издания: 2022
Издательство: Elsevier BV
Местоположение издательства: Netherlands
DOI: 10.1002/1873-3468.14556
Аннотация: The repetitive copying of template nucleotides due to transcriptional slippage has not been reported for RNA-directed RNA polymerases of positive-strand RNA phages. We unexpectedly observed that, with GTP as the only substrate, Qβ replicase, the RNA-directed RNA polymerase of bacteriophage Qβ, synthesizes by transcriptional slippage polyG strands, which on denaturing electrophoresis produce a ladder with at least three clusters of bolder bands. The 15-nt-long G15, the major product of the shortest cluster, is tightly bound by the enzyme but can be released by the ribosomal protein S1, which, as a Qβ replicase subunit, normally promotes the release of a completed transcript. 7-deaza-GTP suppresses the polyG synthesis and abolishes the periodic pattern, suggesting that the N7 atom is needed for the initiation of RNA synthesis and the formation of the structure recognized by protein S1. The results provide new insights into the mechanism of RNA synthesis by the RNA-directed RNA polymerase of a single-stranded RNA phage.
Добавил в систему: Четверин Александр Борисович

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