Chemistry of gene silencing

The mechanism of NAD+-dependent deacetylation reactions

A. A. Sauve, I. Celic, J. Avalos, H. Deng, J. D. Boeke, V. L. Schramm

Research output: Contribution to journalArticle

Abstract

The Sir2 enzyme family is responsible for a newly classified chemical reaction, NAD+-dependent protein deacetylation. New peptide substrates, the reaction mechanism, and the products of the acetyl transfer to NAD+ are described for SIR2. The final products of SIR2 reactions are the deacetylated peptide and the 2′ and 3′ regioisomers of O-acetyl ADP ribose (AADPR), formed through an α-1′-acetyl ADP ribose intermediate and intramolecular transesterification reactions (2′ → 3′). The regioisomers, their anomeric forms, the interconversion rates, and the reaction equilibria were characterized by NMR, HPLC, 18O exchange, and MS methods. The mechanism of acetyl transfer to NAD+ includes (1) ADP ribosylation of the peptide acyl oxygen to form a high-energy O-alkyl amidate intermediate, (2) attack of the 2′-OH group on the amidate to form a 1′,2′ -acyloxonium species, (3) hydrolysis to 2′-AADPR by the attack of water on the carbonyl carbon, and (4) an SIR2-independent transesterification equilibrating the 2′- and 3′-AADPRs. This mechanism is unprecedented in ADP-ribosyl transferase enzymology. The 2′-and 3′-AADPR products are candidate molecules for SIR2-initiated signaling pathways.

Original languageEnglish (US)
Pages (from-to)15456-15463
Number of pages8
JournalBiochemistry®
Volume40
Issue number51
DOIs
StatePublished - Dec 25 2001

Fingerprint

Adenosine Diphosphate Ribose
Gene Silencing
NAD
Genes
Transesterification
Adenosine Diphosphate
Peptides
Transferases
Chemical reactions
Hydrolysis
Carbon
High Pressure Liquid Chromatography
Nuclear magnetic resonance
Oxygen
Molecules
Water
Substrates
Enzymes
Proteins
3-O-acetyl-ADP-ribose

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sauve, A. A., Celic, I., Avalos, J., Deng, H., Boeke, J. D., & Schramm, V. L. (2001). Chemistry of gene silencing: The mechanism of NAD+-dependent deacetylation reactions. Biochemistry®, 40(51), 15456-15463. https://doi.org/10.1021/bi011858j

Chemistry of gene silencing : The mechanism of NAD+-dependent deacetylation reactions. / Sauve, A. A.; Celic, I.; Avalos, J.; Deng, H.; Boeke, J. D.; Schramm, V. L.

In: Biochemistry®, Vol. 40, No. 51, 25.12.2001, p. 15456-15463.

Research output: Contribution to journalArticle

Sauve, AA, Celic, I, Avalos, J, Deng, H, Boeke, JD & Schramm, VL 2001, 'Chemistry of gene silencing: The mechanism of NAD+-dependent deacetylation reactions', Biochemistry®, vol. 40, no. 51, pp. 15456-15463. https://doi.org/10.1021/bi011858j
Sauve AA, Celic I, Avalos J, Deng H, Boeke JD, Schramm VL. Chemistry of gene silencing: The mechanism of NAD+-dependent deacetylation reactions. Biochemistry®. 2001 Dec 25;40(51):15456-15463. https://doi.org/10.1021/bi011858j
Sauve, A. A. ; Celic, I. ; Avalos, J. ; Deng, H. ; Boeke, J. D. ; Schramm, V. L. / Chemistry of gene silencing : The mechanism of NAD+-dependent deacetylation reactions. In: Biochemistry®. 2001 ; Vol. 40, No. 51. pp. 15456-15463.
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