title: Identification and developmental expression of Xenopus laevis SUMO proteases.
abstract: SUMO proteins are small ubiquitin-related modifiers. All SUMOs are synthesized as propeptides that are post-translationally cleaved prior to conjugation. After processing, SUMOs become covalently conjugated to cellular targets through a pathway that is similar to ubiquitination. Ubiquitin like protein proteases/Sentrin specific proteases (Ulp/SENPs) mediate both processing and deconjugation of SUMOs. The action of Ulp/SENPs makes SUMOylation a highly dynamic post-translational modification. To investigate how Ulp/SENPs are regulated in a developmental context, we isolated and characterized all Ulp/SENPs in Xenopus laevis. Xenopus possess homologues of mammalian SENP3, 5, 6 and 7. All of these enzymes reacted with HA-tagged vinyl sulfone derivatives of SUMO-2 (HA-SU2-VS) but not SUMO-1 (HA-SU1-VS), suggesting that they act primarily on SUMO-2 and -3. In contrast, Xenopus possess a single member of the SENP1/SENP2 subfamily of Ulp/SENPs, most closely related to mammalian SENP1. Xenopus SENP1 reacted with HA-SU1-VS and HA-SU2-VS, suggesting that it acts on all SUMO paralogues. We analyzed the mRNA and protein levels for each of the Ulp/SENPs through development; we found that they show distinct patterns of expression that may involve both transcriptional and post-transcriptional regulation. Finally, we have characterized the developmental function of the most abundant Ulp/SENP found within Xenopus eggs, SENP3. Depletion of SENP3 using morpholino antisense oligonucleotides (morpholinos) caused accumulation of high molecular weight SUMO-2/3 conjugated species, defects in developing embryos and changes in the expression of some genes regulated by the transforming growth factor beta (TGF-beta) pathway. These findings collectively indicate that SUMO proteases are both highly regulated and essential for normal development.
authors.affiliation: Laboratory of Gene Regulation and Development, National Institute of Child Health and Human Development, Bethesda, Maryland, United States of America.
authors.forename: Yonggang
authors.initials: Y
authors.lastname: Wang
authors.forename: Debaditya
authors.initials: D
authors.lastname: Mukhopadhyay
authors.forename: Smita
authors.initials: S
authors.lastname: Mathew
authors.forename: Takashi
authors.initials: T
authors.lastname: Hasebe
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Heimeier
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Azuma
authors.forename: Nagamalleswari
authors.initials: N
authors.lastname: Kolli
authors.forename: Yun-Bo
authors.initials: YB
authors.lastname: Shi
authors.forename: Keith D
authors.initials: KD
authors.lastname: Wilkinson
authors.forename: Mary
authors.initials: M
authors.lastname: Dasso
mesh_terms: D000818:Animals; D002457:Cell Extracts; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D010063:Ovum / Q000166:cytology / Q000201:enzymology; D010447:Peptide Hydrolases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity; D016212:Transforming Growth Factor beta / Q000378:metabolism; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics*
chemical_list: D002457:Cell Extracts; D012333:RNA, Messenger; D025841:Small Ubiquitin-Related Modifier Proteins; D016212:Transforming Growth Factor beta; D029867:Xenopus Proteins; D010447:Peptide Hydrolases