A novel mammalian Smt3-specific isopeptidase 1 (SMT3IP1) localized in the nucleolus at interphase.
A novel Smt3-specific isopeptidase, SMT3IP1, was cloned using a yeast two-hybrid screen with Smt3b as bait. The clone, named SMT3IP1 (Smt3-specific isopeptidase 1), which bound to Smt3b but not SUMO-1 in the two-hybrid system, was distantly related to budding yeast Saccharomyces cerevisiae Ulp1, human SENP1 or human SUSP1. The catalytic domains in the C-terminal region were very similar, but the N-terminal region was quite different to other enzymes. The cysteine, histidine and asparatic acid residues in the catalytic domains were conserved. SMT3IP1 expressed by the baculovirus-expression system had the ability to cleave SUMO-1 or Smt3b from SUMO-1/RanGAP1 or Smt3b/RanGAP1 conjugates, respectively, and the activity was a little stronger towards the Smt3b conjugate than towards the SUMO-1 conjugate. Furthermore, the enzyme bound more strongly to Smt3a and Smt3b than to SUMO-1 in vitro. The enzyme did not cleave Nedd8 from Nedd8/cullin-1. Nor did it cleave ubiquitin from ubiquitinated p53. SMT3IP1 was localized almost exclusively at the nucleolus during interphase. The N-terminal sequence was responsible for the nucleolar localization of this enzyme. Whether SMT3IP1 functions in the nucleolus or just stays there before it functions in the nucleus, as shown in the case of CDC14 phosphatase, remains to be elucidated.
Nishida
T
T
School of Life Science, Tokyo University of Parmacy and Life Science, Japan.
Tanaka
H
H
Yasuda
H
H
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002466:Cell Nucleolus / Q000201:enzymology*; D017124:Conserved Sequence; D003546:Cysteine Endopeptidases; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D051379:Mice; D008969:Molecular Sequence Data; D000075687:NEDD8 Protein; D010447:Peptide Hydrolases; D025842:SUMO-1 Protein; D016415:Sequence Alignment; D017384:Sequence Deletion / Q000235:genetics; D025841:Small Ubiquitin-Related Modifier Proteins; D013379:Substrate Specificity; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014452:Ubiquitins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; C513172:Nedd8 protein, mouse; D025842:SUMO-1 Protein; C117909:SUMO2 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; C117908:Sumo3 protein, mouse; D016159:Tumor Suppressor Protein p53; D014452:Ubiquitins; D010450:Endopeptidases; D010447:Peptide Hydrolases; C416522:Smt3-specific isopeptidase 1; D003546:Cysteine Endopeptidases; C492190:Senp3 protein, mouse