Rolling Circle RNA Synthesis Catalysed by RNA

TY - JOUR

T1 - Rolling Circle RNA Synthesis Catalysed by RNA

AU - Kristoffersen, Emil Laust

AU - Burman, Matthew

AU - Noy, Agnes

AU - Holliger, Philipp

N1 - Publisher Copyright: © 2022, eLife Sciences Publications Ltd. All rights reserved.

PY - 2022/2

Y1 - 2022/2

N2 - RNA-catalysed RNA replication is widely considered a key step in the emergence of life’s first genetic system. However, RNA replication can be impeded by the extraordinary stability of duplex RNA products, which must be dissociated for re-initiation of the next replication cycle. Here we have explored rolling circle synthesis (RCS) as a potential solution to this strand separation problem. We observe sustained RCS by a triplet polymerase ribozyme beyond full-length circle synthesis with strand displacement yielding concatemeric RNA products. Furthermore, we show RCS of a circular Hammerhead ribozyme capable of selfcleavage and re-circularisation. Thus, all steps of a viroid-like RNA replication pathway can be catalysed by RNA alone. Finally, we explore potential RCS mechanisms by molecular dynamics simulations, which indicate a progressive build-up of conformational strain upon RCS with destabilisation of nascent strand 5’-and 3’-ends. Our results have implications for the emergence of RNA replication and for understanding the potential of RNA to support complex genetic processes.

AB - RNA-catalysed RNA replication is widely considered a key step in the emergence of life’s first genetic system. However, RNA replication can be impeded by the extraordinary stability of duplex RNA products, which must be dissociated for re-initiation of the next replication cycle. Here we have explored rolling circle synthesis (RCS) as a potential solution to this strand separation problem. We observe sustained RCS by a triplet polymerase ribozyme beyond full-length circle synthesis with strand displacement yielding concatemeric RNA products. Furthermore, we show RCS of a circular Hammerhead ribozyme capable of selfcleavage and re-circularisation. Thus, all steps of a viroid-like RNA replication pathway can be catalysed by RNA alone. Finally, we explore potential RCS mechanisms by molecular dynamics simulations, which indicate a progressive build-up of conformational strain upon RCS with destabilisation of nascent strand 5’-and 3’-ends. Our results have implications for the emergence of RNA replication and for understanding the potential of RNA to support complex genetic processes.

KW - Circular nucleic acids

KW - Circular RNA

KW - Hammerhead

KW - Origin of life

KW - Persistence length

KW - Ribozymes

KW - RNA molecular dynamics simulation

KW - Rolling circle

KW - Rolling circle synthesis

UR - http://www.scopus.com/inward/record.url?scp=85125572248&partnerID=8YFLogxK

U2 - 10.7554/ELIFE.75186

DO - 10.7554/ELIFE.75186

M3 - Journal article

C2 - 35108196

AN - SCOPUS:85125572248

SN - 2050-084X

VL - 11

JO - eLife

JF - eLife

M1 - e75186

ER -