BACKGROUND AND PURPOSE: Small ubiquitin-like modifier (SUMO) conjugation is a post-translational modification associated with many human diseases. Characterization of the SUMO-modified proteome is pivotal to define the mechanistic link between SUMO conjugation and such diseases. This is particularly evident for SUMO2/3 conjugation, which is massively activated after brain ischemia/stroke, and is believed to be a protective response. The purpose of this study was to perform a comprehensive analysis of the SUMO3-modified proteome regulated by brain ischemia using a novel SUMO transgenic mouse. METHODS: To enable SUMO proteomics analysis in vivo, we generated transgenic mice conditionally expressing tagged SUMO1-3 paralogues. Transgenic mice were subjected to 10 minutes forebrain ischemia and 1 hour of reperfusion. SUMO3-conjugated proteins were enriched by anti-FLAG affinity purification and analyzed by liquid chromatography-tandem mass spectrometry. RESULTS: Characterization of SUMO transgenic mice demonstrated that all 3 tagged SUMO paralogues were functionally active, and expression of exogenous SUMOs did not modify the endogenous SUMOylation machinery. Proteomics analysis identified 112 putative SUMO3 substrates of which 91 candidates were more abundant in the ischemia group than the sham group. Data analysis revealed processes/pathways with putative neuroprotective functions, including glucocorticoid receptor signaling, RNA processing, and SUMOylation-dependent ubiquitin conjugation. CONCLUSIONS: The identified proteins/pathways modulated by SUMOylation could be the key to understand the mechanisms linking SUMOylation to neuroprotection, and thus provide new promising targets for therapeutic interventions. The new transgenic mouse will be an invaluable platform for analyzing the SUMO-modified proteome in models of human disorders and thereby help to mechanistically link SUMOylation to the pathological processes.
BACKGROUND AND PURPOSE: Small ubiquitin-like modifier (SUMO) conjugation is a post-translational modification associated with many human diseases. Characterization of the SUMO-modified proteome is pivotal to define the mechanistic link between SUMO conjugation and such diseases. This is particularly evident for SUMO2/3 conjugation, which is massively activated after brain ischemia/stroke, and is believed to be a protective response. The purpose of this study was to perform a comprehensive analysis of the SUMO3-modified proteome regulated by brain ischemia using a novel SUMO transgenicmouse. METHODS: To enable SUMO proteomics analysis in vivo, we generated transgenic mice conditionally expressing tagged SUMO1-3 paralogues. Transgenic mice were subjected to 10 minutes forebrain ischemia and 1 hour of reperfusion. SUMO3-conjugated proteins were enriched by anti-FLAG affinity purification and analyzed by liquid chromatography-tandem mass spectrometry. RESULTS: Characterization of SUMO transgenic mice demonstrated that all 3 tagged SUMO paralogues were functionally active, and expression of exogenous SUMOs did not modify the endogenous SUMOylation machinery. Proteomics analysis identified 112 putative SUMO3 substrates of which 91 candidates were more abundant in the ischemia group than the sham group. Data analysis revealed processes/pathways with putative neuroprotective functions, including glucocorticoid receptor signaling, RNA processing, and SUMOylation-dependent ubiquitin conjugation. CONCLUSIONS: The identified proteins/pathways modulated by SUMOylation could be the key to understand the mechanisms linking SUMOylation to neuroprotection, and thus provide new promising targets for therapeutic interventions. The new transgenicmouse will be an invaluable platform for analyzing the SUMO-modified proteome in models of human disorders and thereby help to mechanistically link SUMOylation to the pathological processes.
Authors: Pablo de Felipe; Garry A Luke; Lorraine E Hughes; David Gani; Claire Halpin; Martin D Ryan Journal: Trends Biotechnol Date: 2005-12-27 Impact factor: 19.536
Authors: John Prudden; Stephanie Pebernard; Grazia Raffa; Daniela A Slavin; J Jefferson P Perry; John A Tainer; Clare H McGowan; Michael N Boddy Journal: EMBO J Date: 2007-08-30 Impact factor: 11.598
Authors: Ivan Matic; Martijn van Hagen; Joost Schimmel; Boris Macek; Stephen C Ogg; Michael H Tatham; Ronald T Hay; Angus I Lamond; Matthias Mann; Alfred C O Vertegaal Journal: Mol Cell Proteomics Date: 2007-10-15 Impact factor: 5.911
Authors: Hari S Sharma; Dafin F Muresanu; Jose V Lafuente; Per-Ove Sjöquist; Ranjana Patnaik; Aruna Sharma Journal: Mol Neurobiol Date: 2015-07-01 Impact factor: 5.590
Authors: Shu Yu; Francesca Galeffi; Ramona M Rodriguiz; Zhuoran Wang; Yuntian Shen; Jingjun Lyu; Ran Li; Joshua D Bernstock; Kory R Johnson; Shuai Liu; Huaxin Sheng; Dennis A Turner; William C Wetsel; Wulf Paschen; Wei Yang Journal: FASEB J Date: 2020-09-10 Impact factor: 5.191