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Roles of dimeric intermediates in RNA-catalyzed rolling circle synthesis.

Emil L Kristoffersen1,2, Ewan K S McRae1,3, Niels R Sørensen1

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RNA-catalyzed rolling circle synthesis (RCS) produces diverse RNA nanostructures, including stable dimeric complexes. These dimers can inhibit replication but may have aided early RNA genetics by storing and coordinating replication.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Origin of Life Studies

Background:

  • The RNA world hypothesis posits RNA's central role in early life.
  • RNA polymerase ribozymes catalyze RNA replication, supporting this hypothesis.
  • RNA-catalyzed rolling circle synthesis (RCS) on small circular RNA (scRNA) templates has been recently established.

Purpose of the Study:

  • To structurally and dynamically characterize scRNA replication products and intermediates.
  • To elucidate the formation and properties of RNA nanostructures in RCS.

Main Methods:

  • Cryogenic electron microscopy (cryo-EM) was employed to analyze RCS products and intermediates.
  • Structural analysis focused on resolving RNA nanostructures at high resolution (5.3 Å).

Main Results:

  • RCS replication yields diverse RNA nanostructures, notably a stable, fully hybridized dimeric complex of scRNAs and their complements.
  • Higher-order dimer filaments and assembly intermediates were observed, suggesting a specific assembly mechanism.
  • Dimer complexes inhibit RCS but can be reactivated by additional scRNA templates.

Conclusions:

  • Dimer formation appears to be a general characteristic of RCS replication.
  • These stable RNA dimers may have served as a storage mechanism for replication products in primordial RNA genetic systems.
  • Dimerization could have facilitated coordinated replication across both scRNA template strands.