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Updated: Jul 1, 2026

Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Crystal structures of RNA 3'-terminal phosphate cyclase and its complexes with Mg2+ +ATP, ATP or Mn2+
Satoru Shimizu1, Masanori Ohki, Nami Ohkubo
1Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
Abstract:
RNA 3'-terminal phosphate cyclase (Rtc) is an enzyme related to RNA splicing, in which the 3'-terminal hydroxyl group of a truncated RNA is converted to the 2',3'-cyclic phosphate that is required prior to RNA ligation. This reaction may occur in the following two steps: (i) Rtc + ATP --> Rtc-AMP + Ppi and (ii) RNA-N3' + Rtc-AMP --> RNA-N>p + Rtc + AMP. In order to establish the reaction mechanism, Rtc of Sulfolobus tokodaii, overexpressed in E. coli, was crystallized in the following states, Rtc, Rtc-AMP, Rtc:AMP, Rtc:ATP and Rtc:Mn, and their crystal structures have been determined at 2.25, 2.25, 2.9, 2.4 and 3.2 A resolutions, respectively. Based on these structures, a possible reaction mechanism has been proposed.
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