Author Contributions Conceived and designed the experiments: CF GM. Performed the experiments: CF LM BA AC EVD FC MM PP ME RSP. Analyzed the References 1. Goldberg AL (2003) Protein degradation and protection against misfolded or damaged proteins. Nature 426: 895–899. 2. Kawaguchi Y, Kovacs JJ, McLaurin A, Vance JM, Ito A, et al. (2003) The deacetylase HDAC6 regulates aggresome formation and cell viability in response to misfolded protein stress. Cell 115: 727–738. 3. Iwata A, Riley BE, Johnston JA, Kopito RR (2005) HDAC6 and microtubules are required for autophagic degradation of aggregated huntingtin. J Biol Chem 280: 40282–40292. 4. Pandey UB, Nie Z, Batlevi Y, McCray BA, Ritson GP, et al. (2007) HDAC6 rescues neurodegeneration and provides an essential link between autophagy and the UPS. Nature 447: 859–863. 5. Matthias P, Yoshida M, Khochbin S (2008) HDAC6 a new cellular stress surveillance factor. Cell Cycle 7: 7–10. 6. Kirkin V, Lamark T, Sou YS, Bjorkoy G, Nunn JL, et al. (2009) A role for NBR1 in autophagosomal degradation of ubiquitinated substrates. Mol Cell 33: 505–516. 7. Komatsu M, Ichimura Y (2010) Physiological significance of selective degradation of p62 by autophagy. FEBS Lett 584: 1374–1378. 8. Komatsu M, Waguri S, Koike M, Sou YS, Ueno T, et al. (2007) Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy- deficient mice. Cell 131: 1149–1163. 9. Chen D, Gao F, Li B, Wang H, Xu Y, et al. (2010) Parkin mono-ubiquitinates Bcl-2 and regulates autophagy. J Biol Chem 285: 38214–38223. 10. Chin LS, Olzmann JA, Li L (2010) Parkin-mediated ubiquitin signalling in aggresome formation and autophagy. Biochem Soc Trans 38: 144–149. 11. Hatakeyama S (2011) TRIM proteins and cancer. Nat Rev Cancer 11: 792–804. 12. Meroni G, Diez-Roux G (2005) TRIM/RBCC, a novel class of ‘single protein RING finger’ E3 ubiquitin ligases. Bioessays 27: 1147–1157. 13. Reymond A, Meroni G, Fantozzi A, Merla G, Cairo S, et al. (2001) The tripartite motif family identifies cell compartments. Embo J 20: 2140–2151. 14. Merla G, Ucla C, Guipponi M, Reymond A (2002) Identification of additional transcripts in the Williams-Beuren syndrome critical region. Hum Genet 110: 429–438. 15. Micale L, Fusco C, Augello B, Napolitano LM, Dermitzakis ET, et al. (2008) Williams-Beuren syndrome TRIM50 encodes an E3 ubiquitin ligase. Eur J Hum Genet 16: 1038–1049. 16. Merla G, Brunetti-Pierri N, Micale L, Fusco C (2010) Copy number variants at Williams-Beuren syndrome 7q11.23 region. Hum Genet. 17. Egorov MV, Capestrano M, Vorontsova OA, Di Pentima A, Egorova AV, et al. (2009) Faciogenital dysplasia protein (FGD1) regulates export of cargo proteins from the golgi complex via Cdc42 activation. Mol Biol Cell 20: 2413–2427. 18. Kawaguchi Y, Ito A, Appella E, Yao TP (2006) Charge modification at multiple C-terminal lysine residues regulates p53 oligomerization and its nucleus- cytoplasm trafficking. J Biol Chem 281: 1394–1400. 19. Zhang Y, Kwon S, Yamaguchi T, Cubizolles F, Rousseaux S, et al. (2008) Mice lacking histone deacetylase 6 have hyperacetylated tubulin but are viable and develop normally. Mol Cell Biol 28: 1688–1701. 20. Gal J, Strom AL, Kilty R, Zhang F, Zhu H (2007) p62 accumulates and enhances aggregate formation in model systems of familial amyotrophic lateral sclerosis. J Biol Chem 282: 11068–11077. 21. Babu JR, Geetha T, Wooten MW (2005) Sequestosome 1/p62 shuttles polyubiquitinated tau for proteasomal degradation. J Neurochem 94: 192–203. 22. Seibenhener ML, Geetha T, Wooten MW (2007) Sequestosome 1/p62–more than just a scaffold. FEBS Lett 581: 175–179. 23. Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, et al. (2007) p62/ SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem 282: 24131–24145. TRIM50 Promotes Aggresome Formation and Clearance data: AF GM. Contributed reagents/materials/analysis tools: GD AR. Wrote the paper: GM. Helped write paper: CF LM ME MM EVD BA FC AC AF RSP GD AR PP GM. 24. Lee JY, Koga H, Kawaguchi Y, Tang W, Wong E, et al. (2010) HDAC6 controls autophagosome maturation essential for ubiquitin-selective quality- control autophagy. EMBO J 29: 969–980. 25. Song B, Javanbakht H, Perron M, Park DH, Stremlau M, et al. (2005) Retrovirus restriction by TRIM5alpha variants from Old World and New World primates. J Virol 79: 3930–3937. 26. Wilde IB, Brack M, Winget JM, Mayor T (2011) Proteomic characterization of aggregating proteins after the inhibition of the ubiquitin proteasome system. J Proteome Res 10: 1062–1072. 27. Song C, Xiao Z, Nagashima K, Li CC, Lockett SJ, et al. (2008) The heavy metal cadmium induces valosin-containing protein (VCP)-mediated aggresome formation. Toxicol Appl Pharmacol 228: 351–363. 28. Gozal YM, Duong DM, Gearing M, Cheng D, Hanfelt JJ, et al. (2009) Proteomics analysis reveals novel components in the detergent-insoluble subproteome in Alzheimer’s disease. J Proteome Res 8: 5069–5079. 29. Meierhofer D, Wang X, Huang L, Kaiser P (2008) Quantitative analysis of global ubiquitination in HeLa cells by mass spectrometry. J Proteome Res 7: 4566–4576. 30. Vasilescu J, Smith JC, Ethier M, Figeys D (2005) Proteomic analysis of ubiquitinated proteins from human MCF-7 breast cancer cells by immunoaffi- nity purification and mass spectrometry. J Proteome Res 4: 2192–2200. 31. Chernorudskiy AL, Garcia A, Eremin EV, Shorina AS, Kondratieva EV, et al. (2007) UbiProt: a database of ubiquitylated proteins. BMC Bioinformatics 8: 126. 32. Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, et al. (2005) p62/ SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171: 603–614. 33. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, et al. (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19: 5720–5728. 34. Gamerdinger M, Kaya AM, Wolfrum U, Clement AM, Behl C (2011) BAG3 mediates chaperone-based aggresome-targeting and selective autophagy of misfolded proteins. EMBO Rep 12: 149–156. 35. Johnston JA, Illing ME, Kopito RR (2002) Cytoplasmic dynein/dynactin mediates the assembly of aggresomes. Cell Motil Cytoskeleton 53: 26–38. 36. Levine B, Kroemer G (2008) Autophagy in the pathogenesis of disease. Cell 132: 27–42. 37. Luciani A, Villella VR, Esposito S, Brunetti-Pierri N, Medina D, et al. (2010) Defective CFTR induces aggresome formation and lung inflammation in cystic fibrosis through ROS-mediated autophagy inhibition. Nat Cell Biol. 38. Ichimura Y, Komatsu M (2010) Selective degradation of p62 by autophagy. Semin Immunopathol 32: 431–436. 39. Lee JY, Nagano Y, Taylor JP, Lim KL, Yao TP (2010) Disease-causing mutations in parkin impair mitochondrial ubiquitination, aggregation, and HDAC6-dependent mitophagy. J Cell Biol 189: 671–679. 40. Takashima A (2001) Establishment of fibroblast cultures. Curr Protoc Cell Biol Chapter 2: Unit 2 1. 41. Polishchuk RS, Polishchuk EV, Marra P, Alberti S, Buccione R, et al. (2000) Correlative light-electron microscopy reveals the tubular-saccular ultrastructure of carriers operating between Golgi apparatus and plasma membrane. J Cell Biol 148: 45–58. 42. Merla G, Howald C, Antonarakis SE, Reymond A (2004) The subcellular localization of the ChoRE-binding protein, encoded by the Williams-Beuren syndrome critical region gene 14, is regulated by 14-3-3. Hum Mol Genet 13: 1505–1514. PLoS ONE | www.plosone.org 13 July 2012 | Volume 7 | Issue 7 | e40440