Design, Synthesis, and Properties of Phosphoramidate 2′,5′-Linked Branched RNA

Toward the Rational Design of Inhibitors of the RNA Lariat Debranching Enzyme

Nobuhiro Tago, Adam Katolik, Nathaniel E. Clark, Eric J. Montemayor, Kohji Seio, Mitsuo Sekine, P. John Hart, Masad J. Damha

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Two RNA fragments linked by means of a 2′,5′ phosphodiester bridge (2′ hydroxyl of one fragment connected to the 5′ hydroxyl of the other) constitute a class of nucleic acids known as 2′-5′ branched RNAs (bRNAs). In this report we show that bRNA analogues containing 2′-5′ phosphoramidate linkages (bN-RNAs) inhibit the lariat debranching enzyme, a 2′,5′-phosphodiesterase that has recently been implicated in neurodegenerative diseases associated with aging. bN-RNAs were efficiently generated using automated solid-phase synthesis and suitably protected branchpoint building blocks. Two orthogonally removable groups, namely the 4-monomethoxytrityl (MMTr) group and the fluorenylmethyl-oxycarbonyl (Fmoc) groups, were evaluated as protecting groups of the 2′ amino functionality. The 2′-N-Fmoc methodology was found to successfully produce bN-RNAs on solid-phase oligonucleotide synthesis. The synthesized bN-RNAs resisted hydrolysis by the lariat debranching enzyme (Dbr1) and, in addition, were shown to attenuate the Dbr1-mediated hydrolysis of native bRNA.

Original languageEnglish (US)
Pages (from-to)10108-10118
Number of pages11
JournalJournal of Organic Chemistry
Volume80
Issue number20
DOIs
StatePublished - Oct 16 2015

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RNA
Hydroxyl Radical
Hydrolysis
Neurodegenerative diseases
Type 5 Cyclic Nucleotide Phosphodiesterases
phosphoramidic acid
lariat debranching enzyme
Oligonucleotides
Nucleic Acids
Aging of materials

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Design, Synthesis, and Properties of Phosphoramidate 2′,5′-Linked Branched RNA : Toward the Rational Design of Inhibitors of the RNA Lariat Debranching Enzyme. / Tago, Nobuhiro; Katolik, Adam; Clark, Nathaniel E.; Montemayor, Eric J.; Seio, Kohji; Sekine, Mitsuo; Hart, P. John; Damha, Masad J.

In: Journal of Organic Chemistry, Vol. 80, No. 20, 16.10.2015, p. 10108-10118.

Research output: Contribution to journalArticle

Tago, Nobuhiro ; Katolik, Adam ; Clark, Nathaniel E. ; Montemayor, Eric J. ; Seio, Kohji ; Sekine, Mitsuo ; Hart, P. John ; Damha, Masad J. / Design, Synthesis, and Properties of Phosphoramidate 2′,5′-Linked Branched RNA : Toward the Rational Design of Inhibitors of the RNA Lariat Debranching Enzyme. In: Journal of Organic Chemistry. 2015 ; Vol. 80, No. 20. pp. 10108-10118.
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