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: ジャーナルへの寄稿記事

21 引用 (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.

元の言語英語
ページ(範囲)2417-2421
ページ数5
ジャーナルJournal of the American Chemical Society
122
発行部数11
DOI
出版物ステータス出版済み - 3 22 2000

Fingerprint

RNA Caps
RNA
Cytosine Nucleotides
Eukaryotic Initiation Factor-4E
DNA Primase
Eukaryotic Initiation Factors
Oligonucleotides
Adenosine
Chemical shift
Magnetic Resonance Spectroscopy
Nucleotides
Metabolism
Labeling
Nuclear magnetic resonance spectroscopy
Messenger RNA
DNA
Proteins
Molecules
m7GpppA

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

これを引用

Matsuo, H., Moriguchi, T., Takagi, T., Kusakabe, T., Buratowski, S., Sekine, M., ... Wagner, G. (2000). Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA. Journal of the American Chemical Society, 122(11), 2417-2421. https://doi.org/10.1021/ja9926820

Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA. / Matsuo, Hiroshi; Moriguchi, Tomohisa; Takagi, Toshimitsu; Kusakabe, Takahiro; Buratowski, Stephen; Sekine, Mitsuo; Kyogoku, Yoshimasa; Wagner, Gerhard.

:: Journal of the American Chemical Society, 巻 122, 番号 11, 22.03.2000, p. 2417-2421.

研究成果: ジャーナルへの寄稿記事

Matsuo, H, Moriguchi, T, Takagi, T, Kusakabe, T, Buratowski, S, Sekine, M, Kyogoku, Y & Wagner, G 2000, 'Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA', Journal of the American Chemical Society, 巻. 122, 番号 11, pp. 2417-2421. https://doi.org/10.1021/ja9926820
Matsuo, Hiroshi ; Moriguchi, Tomohisa ; Takagi, Toshimitsu ; Kusakabe, Takahiro ; Buratowski, Stephen ; Sekine, Mitsuo ; Kyogoku, Yoshimasa ; Wagner, Gerhard. / Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA. :: Journal of the American Chemical Society. 2000 ; 巻 122, 番号 11. pp. 2417-2421.
@article{409411bd47a44264b097eb0cc0f65ca3,
title = "Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA",
abstract = "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.",
author = "Hiroshi Matsuo and Tomohisa Moriguchi and Toshimitsu Takagi and Takahiro Kusakabe and Stephen Buratowski and Mitsuo Sekine and Yoshimasa Kyogoku and Gerhard Wagner",
year = "2000",
month = "3",
day = "22",
doi = "10.1021/ja9926820",
language = "English",
volume = "122",
pages = "2417--2421",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "11",

}

TY - JOUR

T1 - Efficient synthesis of 13C,15N-labeled RNA containing the cap structure m7GpppA

AU - Matsuo, Hiroshi

AU - Moriguchi, Tomohisa

AU - Takagi, Toshimitsu

AU - Kusakabe, Takahiro

AU - Buratowski, Stephen

AU - Sekine, Mitsuo

AU - Kyogoku, Yoshimasa

AU - Wagner, Gerhard

PY - 2000/3/22

Y1 - 2000/3/22

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0034701261&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034701261&partnerID=8YFLogxK

U2 - 10.1021/ja9926820

DO - 10.1021/ja9926820

M3 - Article

AN - SCOPUS:0034701261

VL - 122

SP - 2417

EP - 2421

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 11

ER -