4.8 Article

A ribozyme that triphosphorylates RNA 5'-hydroxyl groups

Journal

NUCLEIC ACIDS RESEARCH
Volume 42, Issue 7, Pages 4767-4778

Publisher

OXFORD UNIV PRESS
DOI: 10.1093/nar/gkt1405

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Funding

  1. National Institutes of Health [T32 GM008326]
  2. Hellman Family foundation
  3. National Aeronautics and Space Administration through the Science Mission Directorate, Research Opportunities in Space and Earth Sciences (ROSES), in Astrobiology/Exobiology [NNX13AJ09G]
  4. NASA [NNX13AJ09G]
  5. NASA [472907, NNX13AJ09G] Funding Source: Federal RePORTER

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The RNA world hypothesis describes a stage in the early evolution of life in which RNA served as genome and as the only genome-encoded catalyst. To test whether RNA world organisms could have used cyclic trimetaphosphate as an energy source, we developed an in vitro selection strategy for isolating ribozymes that catalyze the triphosphorylation of RNA 5'-hydroxyl groups with trimetaphosphate. Several active sequences were isolated, and one ribozyme was analyzed in more detail. The ribozyme was truncated to 96 nt, while retaining full activity. It was converted to a trans-format and reacted with rates of 0.16 min(-1) under optimal conditions. The secondary structure appears to contain a four-helical junction motif. This study showed that ribozymes can use trimetaphosphate to triphosphorylate RNA 5'-hydroxyl groups and suggested that RNA world organisms could have used trimetaphosphate as their energy source.

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