A protein-protein interaction map of the Caenorhabditis elegans 26S proteasome - PubMed (original) (raw)

doi: 10.1093/embo-reports/kve184.

P Bello, N Thierry-Mieg, P Vaglio, J Hitti, L Doucette-Stamm, D Thierry-Mieg, J Reboul, S Boulton, A J Walhout, O Coux, M Vidal

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A protein-protein interaction map of the Caenorhabditis elegans 26S proteasome

A Davy et al. EMBO Rep. 2001 Sep.

Abstract

The ubiquitin-proteasome proteolytic pathway is pivotal in most biological processes. Despite a great level of information available for the eukaryotic 26S proteasome-the protease responsible for the degradation of ubiquitylated proteins-several structural and functional questions remain unanswered. To gain more insight into the assembly and function of the metazoan 26S proteasome, a two-hybrid-based protein interaction map was generated using 30 Caenorhabditis elegans proteasome subunits. The results recapitulate interactions reported for other organisms and reveal new potential interactions both within the 19S regulatory complex and between the 19S and 20S subcomplexes. Moreover, novel potential proteasome interactors were identified, including an E3 ubiquitin ligase, transcription factors, chaperone proteins and other proteins not yet functionally annotated. By providing a wealth of novel biological hypotheses, this interaction map constitutes a framework for further analysis of the ubiquitin-proteasome pathway in a multicellular organism amenable to both classical genetics and functional genomics.

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Figures

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Fig. 1. PCR amplification of pORFs. PCR-amplified ORFs corresponding to potential C. elegans pORFs were analyzed by electrophoresis after organizing them by size in order to facilitate the read-out of the results (Reboul et al., 2001). PCRs were considered successful when a single band of the expected size was observed and accordingly, 30 out of 32 pORFs were successfully PCR amplified and cloned. The PCR product of Y110A7A.14 migrated as a longer fragment than its GeneFinder prediction, however this ORF was confirmed by sequencing (see Reboul et al., 2001 for an explanation). Two pORFs (F10G7.8 and F57B9.10) could not be Gateway cloned due to unsuccessful PCRs.

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Fig. 2. Caenorhabditis elegans proteasome interaction mapping: two-hybrid interactions between 26S proteasome subunits. The data shown here was obtained for all pair-wise combinations by compiling both the matrix (DB-pORF/AD-pORF) and the screens (DB-pORF/AD-cDNAs). See Supplementary data for a detailed description of the comprehensive two-hybrid screens.

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Fig. 3. Proteasome subunit/subunit interactions and new potential interactors. Circles and arrows represent proteins and two-hybrid interactions (from DB-X to AD-Y), respectively. (A) Comprehensive two-hybrid protein network of the C. elegans 26S proteasome. Red circles represent proteasome subunits and green circles represent other potential interactors. (B) Two-hybrid interactions between α and β subunits of the 20S. The α and β rings were separated and each subunit colored differently for clarity. (C) A new model for the 26S proteasome. The 19S subunits used in this study were colored in gray and interactions detected between 26S proteasome subunits are shown with red arrows. Novel potential interactors forming clusters with proteasome subunits and proteins previously characterized for which a model of interaction with the proteasome could be envisaged are shown in green.

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References

    1. Bays N.W., Gardner, R.G., Seelig, L.P., Joazeiro, C.A. and Hampton, R.Y. (2001) Hrd1p/Der3p is a membrane-anchored ubiquitin ligase required for ER-associated degradation. Nature Cell Biol., 3, 24–29. - PubMed
    1. Cagney G., Uetz, P. and Fields, S. (2001) Two-hybrid analysis of the Saccharomyces cerevisiae 26S proteasome. Physiolog. Genomics, in press. - PubMed
    1. Ciechanover A., Orian, A. and Schwartz, A.L. (2000) Ubiquitin-mediated proteolysis: biological regulation via destruction. BioEssays, 97, 2497–2502. - PubMed
    1. Costanzo M.C., et al. (2001) YPD, PombePD and WormPD: model organism volumes of the BioKnowledge library, an integrated resource for protein information. Nucleic Acids Res., 29, 75–79. - PMC - PubMed
    1. Durbin R. and Thierry-Mieg, J. (1994) The ACeDB genome database. In Suhai, S. (ed.), Computational Methods in Genome Research. Plenum Press, New York, NY.

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