GTP-dependent RNA 3'-terminal phosphate cyclase from the hyperthermophilic archaeon Pyrococcus furiosus

Asako Sato, Tomoyoshi Soga, Kaori Igarashi, Kanako Takesue, Masaru Tomita, Akio Kanai

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

We discovered that the PF1549 gene in Pyrococcus furiosus encodes a very heat-stable RNA 3'-terminal phosphate cyclase (Pf-Rtc). Although all previously reported Rtc proteins are ATP-dependent enzymes, we found that Pf-Rtc requires GTP for its cyclase activity at 95°C. Low-level activation of the enzyme was also observed in the presence of dGTP but not other dNTPs, indicating that the guanine base is very important for Pf-Rtc activity. We analyzed a series of GTP analogues and found that the conversion from GTP to GMP is important for Pf-Rtc activity and that an excess of GMP inhibits this activity. Gel-shift analysis clearly showed that the RNA-binding activity of Pf-Rtc is totally dependent on the linear form of the 3'-terminal phosphate, with an apparent K d value of 20nm at 95°C. Furthermore, we found that Pf-Rtc may contribute to GTP-dependent RNA ligation activity through the PF0027 protein (a 2'-5' RNA ligase-like protein in P. furiosus). The possible roles of Pf-Rtc and the importance of terminal phosphate structures in RNA are discussed.

Original languageEnglish
Pages (from-to)1190-1199
Number of pages10
JournalGenes to Cells
Volume16
Issue number12
DOIs
Publication statusPublished - 2011 Dec 1

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

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