Acetylation of Werner protein at K1127 and K1117 is important for nuclear trafficking and DNA repair |
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Affiliation: | 1. Department of Life Science and Biotechnology, Jadavpur University, Kolkata 700032, India;2. Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, United States;1. Thyroid Cancer Registry, Institut Jean-Godinot, 51100 Reims, France;2. EA 3757, University of Reims Champagne-Ardenne, Faculty of Medicine, 51095 Reims, France;3. Department of Research and Innovation, Reims Teaching Hospitals, Robert-Debré Hospital, rue du Général-Koenig, 51092 Reims, France;4. Côte-d’Or Breast Cancer Registry, Georges-François-Leclerc Center, 21000 Dijon, France;1. Facultad de Química, Universidad Autónoma de Querétaro, Centro Universitario, Querétaro 76010, Mexico;2. Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus Juriquilla, Querétaro 76230, Mexico;3. Department of Neurobiology and Behavior, University of California, Irvine, CA 92697-4550, USA;1. Department of Mathematics, Presidency University, Kolkata-700073, India;2. Department of Mathematics, Vidyasagar College, Kolkata-700006, India;3. Department of Mathematics, Jadavpur University, Kolkata-700032, India |
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Abstract: | Werner syndrome is a rare autosomal recessive disorder where Werner (WRN) gene is mutated. Being a nucleolar protein, during DNA damage, WRN translocates at the damage site where its catalytic function is required in DNA repair. Several studies have indicated that WRN acetylation may modulate WRN trafficking and catalytic function (Blander et al., 2002; Lozada et al., 2014). Among the six acetylation sites in WRN protein identified by mass-spectrometry analysis (Li et al., 2010) we here explore the role of acetylation sites in C-terminal of WRN (K1127, K1117, K1389, K1413) because the C- terminal domain is the hub for protein- protein interaction and DNA binding activity (Brosh et al. [4]; Muftuoglu et al., 2008; Huang et al., 2006). To explore their functional activity, we created mutations in these sites by changing the acetylation residue lysine (K) to a non-acetylation residue arginine (R) and expressed them in WRN mutant cell lines. We observed that K1127R and K1117R mutants are sensitive to the DNA damaging agents etoposide and mitomycin C and display deficient DNA repair. Importantly, deacetylation of WRN by SIRT1 (Mammalian Sir2) is necessary for restoration of WRN localization at nucleoli after completion of DNA repair. Among all putative acetylation sites, K1127R, K1117R and the double mutant K1127R/K1117R showed significantly delayed re-entry to the nucleolus after damage recovery, even when SIRT1 is overexpressed. These mutants showed partial interaction with SIRT1 compared to WT WRN. Thus, our results suggest that K1127 and K1117 are the major sites of acetylation, necessary for DNA repair. These results elucidate the mechanism by which SIRT1 regulates WRN trafficking via these acetylation sites during DNA damage. |
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Keywords: | Werner gene Werner syndrome SIRT1 DNA damage Acetylation Post translational modification |
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