title: Distinctive properties of Arabidopsis SUMO paralogues support the in vivo predominant role of AtSUMO1/2 isoforms.
abstract: Protein modification by SUMO (small ubiquitin-related modifier) has emerged as an essential regulatory mechanism in eukaryotes. Even though the molecular mechanisms of SUMO conjugation/deconjugation are conserved, the number of SUMO machinery components and their degree of conservation are specific to each organism. In the present paper, we show data contributing to the notion that the four expressed Arabidopsis SUMO paralogues, AtSUMO1, 2, 3 and 5, have functionally diverged to a higher extent than their human orthologues. We have explored the degree of conservation of these paralogues and found that the surfaces involved in E1-activating enzyme recognition, and E2-conjugating enzyme and SIM (SUMO-interacting motif) non-covalent interactions are well conserved in AtSUMO1/2 isoforms, whereas AtSUMO3 shows a lower degree of conservation, and AtSUMO5 is the most divergent isoform. These differences are functionally relevant, since AtSUMO3 and 5 are deficient in establishing E2 non-covalent interactions, which has not been reported for any naturally occurring SUMO orthologue. In addition, AtSUMO3 is less efficiently conjugated than AtSUMO1/2, and AtSUMO5 shows the lowest conjugation level. A mutagenesis analysis revealed that decreases in conjugation rate and thioester-bond formation are the result of the non-conserved residues involved in E1-activating enzyme recognition that are present in AtSUMO3 and 5. The results of the present study support a role for the E1-activating enzyme in SUMO paralogue discrimination, providing a new mechanism to favour conjugation of the essential AtSUMO1/2 paralogues.
authors.affiliation: Department of Molecular Genetics, Center for Research in Agricultural Genomics CSIC-IRTA-UAB, Parc de Recerca UAB, Edifici CRAG, Campus UAB, 08193 Bellaterra Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Castaño-Miquel
authors.forename: Josep
authors.initials: J
authors.lastname: Seguí
authors.forename: L Maria
authors.initials: LM
authors.lastname: Lois
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D025842:SUMO-1 Protein / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D058207:Sumoylation
chemical_list: D029681:Arabidopsis Proteins; C477512:SUM1 protein, Arabidopsis; C477513:SUM2 protein, Arabidopsis; D025842:SUMO-1 Protein