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Mutations in SIRT2 deacetylase which regulate enzymatic activity but not its interaction with HDAC6 and tubulin

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Abstract

Human SIRT2 is a cytoplasmic NAD-dependent deacetylase implicated in the mitotic regulation of microtubule dynamics by its association with the class II histone deacetylase 6 (HDAC6). We have previously reported that SIRT2 is multiply phosphorylated in a cell cycle dependent pattern. Here, we demonstrate that HDAC6 binds to both phosphorylated and unphosphorylated forms of SIRT2 and that tubulin binds only to the SIRT2–HDAC6 complex. Tubulin does not bind to either HDAC6 or SIRT2 individually. In addition, we show that replacement of specific serines with alanines in either isoform of SIRT2 regulates its enzymatic activity. We also found that overexpression of isoform2 was deleterious to cell survival. SIRT2 was found to be phosphorylated at serines 368 and 372, outside the conserved core domain of the Sir2 protein family. Double replacement of S368A and S372A reduced SIRT2 deacetylase activity by 44% compared to wildtype activity. Replacements of other serine, threonine, and tyrosine residues, which did not alter the phosphorylation pattern, had varying effects on SIRT2 deacetylase activity but no effect on tubulin/HDAC6 binding.

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Acknowledgements

We would like to thank Anton-Scott Goustin for assistance in the design of some of the primers for the mutagenesis of SIRT2 as well as for many helpful discussions and Megan Stojcevski for her computer graphic skills. This work was supported by the Barbara and Fred Erb Endowed Chair in Cancer Genetics to M.A.T.; funds from the Karmanos Cancer Institute (P30CA022453); specifically the Applied Genomics Core and Biostatistics Core of the Karmanos Cancer Institute.

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Authors and Affiliations

  1. Program in Molecular Biology and Human Genetics, Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine, 110 East Warren, Detroit, MI, 48201, USA

    Fatimah Nahhas, Sylvia C. Dryden & Michael A. Tainsky

  2. Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI, USA

    Michael A. Tainsky

  3. Biostatistics Core, Barbara Ann Karmanos Cancer Institute, Detroit, MI, 48201, USA

    Judith Abrams

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  1. Fatimah Nahhas
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  4. Michael A. Tainsky
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Correspondence to Michael A. Tainsky.

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Nahhas, F., Dryden, S.C., Abrams, J. et al. Mutations in SIRT2 deacetylase which regulate enzymatic activity but not its interaction with HDAC6 and tubulin. Mol Cell Biochem 303, 221–230 (2007). https://doi.org/10.1007/s11010-007-9478-6

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