Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA

Hiroshi Matsuo, Tomohisa Moriguchi, Toshimitsu Takagi, Takahiro Kusakabe, Stephen Buratowski, Mitsuo Sekine, Yoshimasa Kyogoku, Gerhard Wagner

研究成果: Contribution to journalArticle査読

21 被引用数 (Scopus)

抄録

Studying the mechanism by which the cap structure performs its numerous biological roles in mRNA metabolism by NMR spectroscopy requires the large- scale production of isotopically-labeled capped RNA. We present an efficient method for production of short RNA containing the cap structure m7GpppA, by combining chemical synthesis with enzymatic synthesis using a DNA primase. This method was employed to synthesize three capped RNA molecules, m7Gppp([13C,15N])A, m7Gppp([13C, 15N])ACC, and m7GpppA([13C,15N])C([13C,15N])C, that contain selective 13C,15N- labeled adenosine or cytosine nucleotides. This selective labeling technique enabled us to obtain the chemical shift assignments of these oligonucleotides in complex with the eukaryotic translation initiation factor 4E (eIF4E). Furthermore, we were able to observed intermolecular NOE interactions between the RNA and protein.

本文言語英語
ページ(範囲)2417-2421
ページ数5
ジャーナルJournal of the American Chemical Society
122
11
DOI
出版ステータス出版済み - 3 22 2000

All Science Journal Classification (ASJC) codes

  • 触媒
  • 化学 (全般)
  • 生化学
  • コロイド化学および表面化学

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