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Rolling circle RNA synthesis catalyzed by RNA.

Emil Laust Kristoffersen1, Matthew Burman2, Agnes Noy2

  • 1MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, United Kingdom.

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|February 2, 2022
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RNA replication, crucial for early life, faces challenges with stable RNA duplexes. Rolling circle synthesis (RCS) by ribozymes offers a solution, enabling sustained RNA replication and complex genetic processes.

Keywords:
biochemistrychemical biologycircular RNAnoneorigin of liferibozymesrolling circle

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Area of Science:

  • Origin of Life
  • RNA Biology
  • Biochemistry

Background:

  • RNA-catalyzed RNA replication is theorized as a foundational step in the emergence of life.
  • The stability of double-stranded RNA products hinders the continuous RNA replication cycle.
  • Strand separation is a critical bottleneck for efficient RNA replication.

Purpose of the Study:

  • To investigate rolling circle synthesis (RCS) as a mechanism to overcome RNA duplex stability issues.
  • To demonstrate RNA-only catalysis of a complete viroid-like replication pathway.
  • To explore the molecular mechanisms underlying RCS in RNA replication.

Main Methods:

  • Utilized a triplet polymerase ribozyme for rolling circle synthesis (RCS) experiments.
  • Investigated RCS of a circular Hammerhead ribozyme with self-cleavage and re-circularization capabilities.
  • Employed molecular dynamics simulations to analyze RCS mechanisms and conformational strain.

Main Results:

  • Observed sustained RCS by a ribozyme, producing concatemeric RNA products through strand displacement.
  • Demonstrated RNA-catalyzed self-cleavage and re-circularization of a Hammerhead ribozyme, completing a replication cycle.
  • Molecular dynamics simulations revealed conformational strain accumulation during RCS, destabilizing RNA ends.

Conclusions:

  • RNA alone can catalyze all steps of a viroid-like RNA replication pathway.
  • RCS offers a viable mechanism for overcoming strand separation limitations in early RNA replication.
  • These findings provide insights into the emergence of RNA replication and the potential of RNA for complex genetic systems.