Tissue-specificity, functional characterization and subcellular localization of a rat ubiquitin-specific processing protease, UBP109, whose mRNA expression is developmentally regulated

Kyung Chan Park, Eun Jung Choi, Sang Won Min, Sung Soo Chung, Hyun Kim, Toshiaki Suzuki, Keiji Tanaka, Chin Ha Chung

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

A cDNA encoding an ubiquitin-specific processing protease, UBP109, in rat skeletal muscle was cloned and its product was characterized. Northern analysis revealed that UBP109 mRNA is highly expressed in testis and spleen, compared with other tissues. Furthermore, in situ hybridization showed that the level of UBP109 mRNA in liver, spinal cord and brain dramatically changed during embryonic development, indicating that the expression of UBP109 mRNA is developmentally regulated. UBP109 was expressed in Escherichia coli and purified to apparent homogeneity using a 123I-labelled ubiquitin-peptide fusion as a substrate. The purified enzyme cleaved at the C-terminus of the ubiquitin moiety in natural and engineered fusions irrespective of their sizes. UBP109 also released free ubiquitin from poly-His-tagged penta-ubiquitin. Moreover, it released free ubiquitin from poly-ubiquitinated protein conjugates of rabbit reticulocytes. In addition, UBP109 localized to both the cytoplasm and the nucleus and, among three putative nuclear localization sequences, only the one located near the C-terminus is responsible for nuclear localization. These results suggest that UBP109 may play an important role in generation of free ubiquitin from its precursors and its recycling from poly-ubiquitinated protein conjugates, and hence in regulation of ubiquitin-mediated cellular processes, particularly related to embryonic development.

Original languageEnglish
Pages (from-to)443-453
Number of pages11
JournalBiochemical Journal
Volume349
Issue number2
DOIs
Publication statusPublished - 2000 Jul 15

Keywords

  • De-ubiquitinating enzyme
  • Embryonic development
  • Nuclear localization
  • UBP109
  • hUSP15

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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