Side chain specificity of ADP-ribosylation by a sirtuin

Kamau Fahie, Po Hu, Stephen Swatkoski, Robert J. Cotter, Yingkai Zhang, Cynthia Wolberger

Research output: Contribution to journalArticle

Abstract

Endogenous mono-ADP-ribosylation in eukaryotes is involved in regulating protein synthesis, signal transduction, cytoskeletal integrity, and cell proliferation, although few cellular ADP-ribosyltransferases have been identified. The sirtuins constitute a highly conserved family of protein deacetylases, and several family members have also been reported to perform protein ADP-ribosylation. We characterized the ADP-ribosylation reaction of the nuclear sirtuin homolog Trypanosoma brucei SIR2-related protein 1 (TbSIR2RP1) on both acetylated and unacetylated substrates. We demonstrated that an acetylated substrate is not required for ADP-ribosylation to occur, indicating that the reaction performed by TbSIR2RP1 is a genuine enzymatic reaction and not a side reaction of deacetylation. Biochemical and MS data showed that arginine is the major ADP-ribose acceptor for unacetylated substrates, whereas arginine does not appear to be the major ADP-ribose acceptor in reactions with acetylated histone H1.1. We performed combined ab initio quantum mechanical\molecular mechanical molecular dynamics simulations, which indicated that sirtuin ADP-ribosylation at arginine is energetically feasible, and involves a concerted mechanism with a highly dissociative transition state. In comparison with the corresponding nicotinamide cleavage in the deacetylation reaction, the simulations suggest that sirtuin ADP-ribosylation would be several orders slower but less sensitive to nicotinamide inhibition, which is consistent with experimental results. These results suggest that TbSIR2RP1 can perform ADP-ribosylation using two distinct mechanisms, depending on whether or not the substrate is acetylated.

Original languageEnglish (US)
Pages (from-to)7159-7176
Number of pages18
JournalFEBS Journal
Volume276
Issue number23
DOIs
StatePublished - Dec 2009

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Adenosine Diphosphate
Trypanosoma brucei brucei
Adenosine Diphosphate Ribose
Arginine
Proteins
Niacinamide
Substrates
Sirtuins
ADP Ribose Transferases
Signal transduction
Cell proliferation
Molecular Dynamics Simulation
Eukaryota
Histones
Molecular dynamics
Signal Transduction
Cell Proliferation
Computer simulation

Keywords

  • ADP-ribose acceptor
  • ADP-ribosyltransferase
  • Mono-ADP-ribosylation
  • Sirtuin
  • TbSIR2

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Fahie, K., Hu, P., Swatkoski, S., Cotter, R. J., Zhang, Y., & Wolberger, C. (2009). Side chain specificity of ADP-ribosylation by a sirtuin. FEBS Journal, 276(23), 7159-7176. https://doi.org/10.1111/j.1742-4658.2009.07427.x

Side chain specificity of ADP-ribosylation by a sirtuin. / Fahie, Kamau; Hu, Po; Swatkoski, Stephen; Cotter, Robert J.; Zhang, Yingkai; Wolberger, Cynthia.

In: FEBS Journal, Vol. 276, No. 23, 12.2009, p. 7159-7176.

Research output: Contribution to journalArticle

Fahie, K, Hu, P, Swatkoski, S, Cotter, RJ, Zhang, Y & Wolberger, C 2009, 'Side chain specificity of ADP-ribosylation by a sirtuin', FEBS Journal, vol. 276, no. 23, pp. 7159-7176. https://doi.org/10.1111/j.1742-4658.2009.07427.x
Fahie K, Hu P, Swatkoski S, Cotter RJ, Zhang Y, Wolberger C. Side chain specificity of ADP-ribosylation by a sirtuin. FEBS Journal. 2009 Dec;276(23):7159-7176. https://doi.org/10.1111/j.1742-4658.2009.07427.x
Fahie, Kamau ; Hu, Po ; Swatkoski, Stephen ; Cotter, Robert J. ; Zhang, Yingkai ; Wolberger, Cynthia. / Side chain specificity of ADP-ribosylation by a sirtuin. In: FEBS Journal. 2009 ; Vol. 276, No. 23. pp. 7159-7176.
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