Unknown

Dataset Information

0

Backtracking behavior in viral RNA-dependent RNA polymerase provides the basis for a second initiation site.


ABSTRACT: Transcription in RNA viruses is highly dynamic, with a variety of pauses interrupting nucleotide addition by RNA-dependent RNA polymerase (RdRp). For example, rare but lengthy pauses (>20 s) have been linked to backtracking for viral single-subunit RdRps. However, while such backtracking has been well characterized for multi-subunit RNA polymerases (RNAPs) from bacteria and yeast, little is known about the details of viral RdRp backtracking and its biological roles. Using high-throughput magnetic tweezers, we quantify the backtracking by RdRp from the double-stranded (ds) RNA bacteriophage ?6, a model system for RdRps. We characterize the probability of entering long backtracks as a function of force and propose a model in which the bias toward backtracking is determined by the base paring at the dsRNA fork. We further discover that extensive backtracking provides access to a new 3'-end that allows for the de novo initiation of a second RdRp. This previously unidentified behavior provides a new mechanism for rapid RNA synthesis using coupled RdRps and hints at a possible regulatory pathway for gene expression during viral RNA transcription.

SUBMITTER: Dulin D 

PROVIDER: S-EPMC4666362 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

altmetric image

Publications

Backtracking behavior in viral RNA-dependent RNA polymerase provides the basis for a second initiation site.

Dulin David D   Vilfan Igor D ID   Berghuis Bojk A BA   Poranen Minna M MM   Depken Martin M   Dekker Nynke H NH  

Nucleic acids research 20151022 21


Transcription in RNA viruses is highly dynamic, with a variety of pauses interrupting nucleotide addition by RNA-dependent RNA polymerase (RdRp). For example, rare but lengthy pauses (>20 s) have been linked to backtracking for viral single-subunit RdRps. However, while such backtracking has been well characterized for multi-subunit RNA polymerases (RNAPs) from bacteria and yeast, little is known about the details of viral RdRp backtracking and its biological roles. Using high-throughput magneti  ...[more]

Similar Datasets

| S-EPMC7611809 | biostudies-literature
| S-EPMC10245719 | biostudies-literature
| S-EPMC4148879 | biostudies-literature
| S-EPMC10790778 | biostudies-literature
| S-EPMC10197661 | biostudies-literature
| S-EPMC5921838 | biostudies-literature
| S-EPMC4858960 | biostudies-literature
| S-EPMC4948327 | biostudies-literature
| S-EPMC3160732 | biostudies-literature
| S-EPMC3197207 | biostudies-literature