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SUMOylation Inhibition Mediated by Disruption of SUMO El-E2 Interactions Confers Plant Susceptibility to Necrotrophic Fungal Pathogens

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Molecular Plant 2017年第5期10卷 709-720页

作者： Laura Castano-Miquel Abraham Mas Ines Teixeira josep segui Anna Perearnau Bhagyasree N. Thampi Arnaldo L. Schapire Natalia Rodrigo Gaelle La Verde Silvia Manrique Maria Coca L. Maria Lois Center for Research in Agricultural Genomics - CRAG Edifici CRAG-Campus UAB Bellaterra (Cerdanyola del Vall~s) 08193 Barcelona Spain

Protein modification by SUMO modulates essential biological processes in eukaryotes. SUMOylation is facilitated by sequential action of the El-activating, E2-conjugating, and E3-1igase enzymes. In plants, SUMO regulates plant development and stress responses, which are key determinants in agricultural productivity. To generate additional tools for advancing our knowledge about the SUMO biology, we have developed a strategy for inhibiting in vivo SUMO conjugation based on disruption of SUMO El-E2 interactions through expression of E1 SAE2uFDct domain. Targeted mutagenesis and phylogenetic analyses revealed that this inhibition involves a short motif in SAE2uFDct highly divergent across kingdoms. Transgenic plants expressing the SAE2UFDCt domain displayed dose-dependent inhibition of SUMO conjugation, and have revealed the existence of a post-transcriptional mechanism that regulates SUMO E2 conjugating enzyme levels. Interestingly, these transgenic plants displayed increased suscep- tibility to necrotrophic fungal infections by Botrytis cinerea and Plectosphaerella cucumerina. Early after fungal inoculation, host SUMO conjugation was post-transcriptionally downregulated, suggesting that targeting SUMOylation machinery could constitute a novel mechanism for fungal pathogenicity. These findings support the role of SUMOylation as a mechanism involved in plant protection from environmental stresses. In addition, the strategy for inhibiting SUMO conjugation in vivo described in this study might be applicable in important crop plants and other non-plant organisms regardless of their genetic complexity.

关键词： SUMQ inhibition development flowering necrotrophic fungi El-activating enzyme El-E2interaction disruption

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Diversification of SUMO-Activating Enzyme in Arabidopsis： Implications in SUMO Conjugation

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Molecular Plant 2013年第5期6卷 1646-1660页

作者： Laura Castano-Miquel josep segui Silvia Manrique Ines Teixeira Lorenzo Carretero-Paulet Felix Atencio L. Maria Lois Center for Research in Agricultural Genomics CRAG (CSIC-IRTA-UAB-UB) Edifici CRAG-Campus UAB Bellaterra (Cerdanyola del Valles) 08193 Barcelona Spain Department of Biological Sciences SUNY-University at Buffalo North Campus 109 Cooke Hall Buffalo NY 14260 USA

Sumoylation is an essential posttranslational modification that participates in many biological processes including stress responses. However, little is known about the mechanisms that control Small Ubiquitin-like MOdifier （SUMO） conjugation in vivo. We have evaluated the regulatory role of the heterodimeric E1 activating enzyme, which catalyzes the first step in SUMO conjugation. We have established that the E1 large SAE2 and small SAE1 subunits are encoded by one and three genes, respectively, in the Arabidopsis genome. The three paralogs genes SAEla, SAElbl, and SAElb2 are the result of two independent duplication events. Since SAElbl and SAElb2 correspond to two identical cop- ies, only two E1 small subunit isoforms are present in vivo： SAEla and SAElb. The E1 heterodimer nuclear localization is modulated by the C-terminal tail of the SAE2 subunit. In vitro, SUMO conjugation rate is dependent on the SAE1 isoform contained in the E1 holoenzyme and our results suggest that downstream steps to SUMO-E1 thioester bond formation are affected. In vivo, SAEla isoform deletion in T-DNA insertion mutant plants conferred sumoylation defects upon abi- otic stress, consistent with a sumoylation defective phenotype. Our results support previous data pointing to a regula- tory role of the E1 activating enzyme during SUMO conjugation and provide a novel mechanism to control sumoylation in vivo by diversification of the E1 small subunit.

关键词： E1 activating enzyme conjugation rate subcellular localization regulation abiotic stress gene duplication.

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