Proximity-induced activation of human Cdc34 through heterologous dimerization Journal Article
| Authors: | Gazdoiu, S.; Yamoah, K.; Wu, K.; Escalante, C. R.; Tappin, I.; Bermudez, V.; Aggarwal, A. K.; Hurwitz, J.; Pan, Z. Q. |
| Article Title: | Proximity-induced activation of human Cdc34 through heterologous dimerization |
| Abstract: | Cdc34 is an E2-conjugating enzyme required for catalyzing the polyubiquitination reaction mediated by the Skp1-CUL1-F-box (SCF) protein E3 ubiquiiin (Ub) ligase. Here, we show that the activity of human Cdc34 in the Ub-Ub ligation reaction was enhanced dramatically by SCF's core Ub ligase module, composed of a heterodimeric complex formed by the ROC1 RING finger protein and the CUL1 C terminus that contains a NeddS moiety covalently conjugated at K720. Unexpectedly, we found that N-terminal fusion of a GST moiety to human Cdc34 generated dimeric GST-Cdc34 that was constitutively active in supporting the assembly of K48-linked polyUb chains independently of SCF. Furthermore, fusion of a FK506-binding protein (FKBP) to the N terminus of human Cdc34 yielded FKBP-Cdc34 that was induced to form a dimer upon treatment with the chemical inducer AP20187. The AP20187-induced dimeric form of FKBP-Cdc34 was substantially more active than the monomer in catalyzing Ub-Ub ligation. Thus, juxtaposition of human Cdc34 activates its catalytic capability, suggesting that the SCF-mediated polyubiquitination reaction may require the conversion of Cdc34 from an inactive monomer to a highly active dimeric form. © 2005 by The National Academy of Sciences of the USA. |
| Keywords: | unclassified drug; ubiquitin; cell cycle proteins; enzyme activation; enzyme activity; ubiquitination; recombinant fusion proteins; carrier proteins; glutathione transferase; dimerization; tacrolimus; ubiquitin-conjugating enzymes; protein structure, quaternary; polyubiquitin; ubiquitin-protein ligases; enzyme purification; enzyme assay; cullin proteins; enzyme; ubiquitin-protein ligase complexes; scf; fk 506 binding protein; tacrolimus binding proteins; cdc34 protein |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 102 |
| Issue: | 42 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2005-10-18 |
| Start Page: | 15053 |
| End Page: | 15058 |
| Language: | English |
| DOI: | 10.1073/pnas.0507646102 |
| PUBMED: | 16210246 |
| PROVIDER: | scopus |
| PMCID: | PMC1242854 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 24" - "Export Date: 24 October 2012" - "CODEN: PNASA" - "Source: Scopus" |
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