RNA-catalyzed RNA ligation on an external RNA template

Chem Biol. 2002 Mar;9(3):297-307. doi: 10.1016/s1074-5521(02)00110-2.

Abstract

Variants of the hc ligase ribozyme, which catalyzes ligation of the 3' end of an RNA substrate to the 5' end of the ribozyme, were utilized to evolve a ribozyme that catalyzes ligation reactions on an external RNA template. The evolved ribozyme catalyzes the joining of an oligonucleotide 3'-hydroxyl to the 5'-triphosphate of an RNA hairpin molecule. The ribozyme can also utilize various substrate sequences, demonstrating a largely sequence-independent mechanism for substrate recognition. The ribozyme also carries out the ligation of two oligonucleotides that are bound at adjacent positions on a complementary template. Finally, it catalyzes addition of mononucleoside 5'-triphosphates onto the 3' end of an oligonucleotide primer in a template-dependent manner. The development of ribozymes that catalyze polymerase-type reactions contributes to the notion that an RNA world could have existed during the early history of life on Earth.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Directed Molecular Evolution
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Nucleic Acid Conformation
  • RNA Ligase (ATP) / metabolism
  • RNA, Catalytic / chemistry
  • RNA, Catalytic / genetics*
  • RNA, Catalytic / metabolism*
  • Sequence Analysis, RNA
  • Substrate Specificity
  • Templates, Genetic

Substances

  • RNA, Catalytic
  • RNA Ligase (ATP)