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Updated: Mar 9, 2026

Deacetylation Assays to Unravel the Interplay between Sirtuins SIRT2 and Specific Protein-substrates
Published on: February 27, 2016
SIRT7 Is an RNA-Activated Protein Lysine Deacylase
Zhen Tong1, Miao Wang1, Yi Wang2
1Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Cornell University , Ithaca, New York 14853, United States.
Abstract:
Mammalian SIRT7 is a member of the sirtuin family that regulates multiple biological processes including genome stability, metabolic pathways, stress responses, and tumorigenesis. SIRT7 has been shown to be important for ribosome biogenesis and transcriptional regulation. SIRT7 knockout mice exhibit complications associated with fatty liver and increased aging in hematopoietic stem cells. However, the molecular basis for its biological function remains unclear, in part due to the lack of efficient enzymatic activity in vitro. Previously, we have demonstrated that double-stranded DNA could activate SIRT7's deacetylase activity in vitro, allowing it to deacetylate H3K18 in the context of chromatin. Here, we show that RNA can increase the catalytic efficiency of SIRT7 even better and that SIRT7 can remove long chain fatty acyl groups more efficiently than removing acetyl groups. Truncation and mutagenesis studies revealed residues at both the amino and carboxyl termini of SIRT7 that are involved in RNA-binding and important for activity. RNA immunoprecipitation-sequencing (RIP-seq) identified ribosomal RNA (rRNA) as the predominant RNA binding partner of SIRT7. The associated RNA was able to effectively activate the deacetylase and defatty-acylase activities of SIRT7. Knockdown of SIRT7 increased the lysine fatty acylation of several nuclear proteins based on metabolic labeling with an alkyne-tagged fatty acid analog, supporting that the defatty-acylase activity of SIRT7 is physiologically relevant. These findings provide important insights into the biological functions of SIRT7, as well as an improved platform to develop SIRT7 modulators.
Insights
RNA significantly enhances the activity of mammalian SIRT7, a sirtuin protein. This protein deacetylates and defatty-acylates nuclear proteins, impacting genome stability and metabolic pathways.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- SIRT7, a sirtuin family member, regulates genome stability, metabolism, and tumorigenesis.
- Its precise molecular functions are unclear due to limited in vitro enzymatic activity.
- Previous studies showed double-stranded DNA activates SIRT7's deacetylase activity.
Purpose of the Study:
- To investigate the role of RNA in SIRT7 activity.
- To characterize SIRT7's enzymatic activities beyond deacetylation.
- To identify SIRT7's RNA binding partners and functional domains.
Main Methods:
- In vitro enzymatic assays with RNA.
- Truncation and mutagenesis studies of SIRT7.
- RNA immunoprecipitation-sequencing (RIP-seq).
- Metabolic labeling to assess lysine fatty acylation.
Main Results:
- RNA enhances SIRT7's catalytic efficiency more than DNA.
- SIRT7 efficiently removes long-chain fatty acyl groups, not just acetyl groups.
- Ribosomal RNA (rRNA) is the primary RNA binding partner for SIRT7.
- SIRT7 knockdown increases lysine fatty acylation of nuclear proteins.
Conclusions:
- RNA binding, particularly rRNA, is crucial for activating SIRT7's deacetylase and defatty-acylase functions.
- SIRT7's defatty-acylase activity is physiologically relevant.
- These findings elucidate SIRT7's biological roles and offer a basis for developing SIRT7 modulators.
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