Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide.
Calpain proteases drive intracellular signal transduction via specific proteolysis of multiple substrates upon Ca(2+)-induced activation. Recently, dUTPase, an enzyme essential to maintain genomic integrity, was identified as a physiological calpain substrate in Drosophila cells. Here we investigate...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3098232?pdf=render |
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| author | Zoltán Bozóky Gergely Róna Éva Klement Katalin F Medzihradszky Gábor Merényi Beáta G Vértessy Peter Friedrich |
| author_facet | Zoltán Bozóky Gergely Róna Éva Klement Katalin F Medzihradszky Gábor Merényi Beáta G Vértessy Peter Friedrich |
| author_sort | Zoltán Bozóky |
| collection | DOAJ |
| description | Calpain proteases drive intracellular signal transduction via specific proteolysis of multiple substrates upon Ca(2+)-induced activation. Recently, dUTPase, an enzyme essential to maintain genomic integrity, was identified as a physiological calpain substrate in Drosophila cells. Here we investigate the potential structural/functional significance of calpain-activated proteolysis of human dUTPase.Limited proteolysis of human dUTPase by mammalian m-calpain was investigated in the presence and absence of cognate ligands of either calpain or dUTPase. Significant proteolysis was observed only in the presence of Ca(II) ions, inducing calpain action. The presence or absence of the dUTP-analogue α,β-imido-dUTP did not show any effect on Ca(2+)-calpain-induced cleavage of human dUTPase. The catalytic rate constant of dUTPase was unaffected by calpain cleavage. Gel electrophoretic analysis showed that Ca(2+)-calpain-induced cleavage of human dUTPase resulted in several distinctly observable dUTPase fragments. Mass spectrometric identification of the calpain-cleaved fragments identified three calpain cleavage sites (between residues (4)SE(5); (7)TP(8); and (31)LS(32)). The cleavage between the (31)LS(32) peptide bond specifically removes the flexible N-terminal nuclear localization signal, indispensable for cognate localization.Results argue for a mechanism where Ca(2+)-calpain may regulate nuclear availability and degradation of dUTPase. |
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| id | doaj.art-338e8e904b4f4aca9e294f05fac91ec5 |
| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-12-13T18:22:35Z |
| publishDate | 2011-01-01 |
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| series | PLoS ONE |
| spelling | doaj.art-338e8e904b4f4aca9e294f05fac91ec52022-12-21T23:35:41ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0165e1954610.1371/journal.pone.0019546Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide.Zoltán BozókyGergely RónaÉva KlementKatalin F MedzihradszkyGábor MerényiBeáta G VértessyPeter FriedrichCalpain proteases drive intracellular signal transduction via specific proteolysis of multiple substrates upon Ca(2+)-induced activation. Recently, dUTPase, an enzyme essential to maintain genomic integrity, was identified as a physiological calpain substrate in Drosophila cells. Here we investigate the potential structural/functional significance of calpain-activated proteolysis of human dUTPase.Limited proteolysis of human dUTPase by mammalian m-calpain was investigated in the presence and absence of cognate ligands of either calpain or dUTPase. Significant proteolysis was observed only in the presence of Ca(II) ions, inducing calpain action. The presence or absence of the dUTP-analogue α,β-imido-dUTP did not show any effect on Ca(2+)-calpain-induced cleavage of human dUTPase. The catalytic rate constant of dUTPase was unaffected by calpain cleavage. Gel electrophoretic analysis showed that Ca(2+)-calpain-induced cleavage of human dUTPase resulted in several distinctly observable dUTPase fragments. Mass spectrometric identification of the calpain-cleaved fragments identified three calpain cleavage sites (between residues (4)SE(5); (7)TP(8); and (31)LS(32)). The cleavage between the (31)LS(32) peptide bond specifically removes the flexible N-terminal nuclear localization signal, indispensable for cognate localization.Results argue for a mechanism where Ca(2+)-calpain may regulate nuclear availability and degradation of dUTPase.http://europepmc.org/articles/PMC3098232?pdf=render |
| spellingShingle | Zoltán Bozóky Gergely Róna Éva Klement Katalin F Medzihradszky Gábor Merényi Beáta G Vértessy Peter Friedrich Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. PLoS ONE |
| title | Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. |
| title_full | Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. |
| title_fullStr | Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. |
| title_full_unstemmed | Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. |
| title_short | Calpain-catalyzed proteolysis of human dUTPase specifically removes the nuclear localization signal peptide. |
| title_sort | calpain catalyzed proteolysis of human dutpase specifically removes the nuclear localization signal peptide |
| url | http://europepmc.org/articles/PMC3098232?pdf=render |
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