The de novo synthesis of ubiquitin: Identification of deubiquitinases acting on ubiquitin precursors
| Autor: | Jorge E. Azevedo, Manuel P. Pinto, Cláudia P. Grou, Andreia V. Mendes, Pedro Domingues |
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| Přispěvatelé: | Instituto de Investigação e Inovação em Saúde |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Ribosomal Proteins
Liver/metabolism Ubiquitins/metabolism Ubiquitin-Activating Enzymes Ubiquitin-conjugating enzyme Article Deubiquitinating enzyme Mice Ubiquitin Endopeptidases/metabolism Ubiquitin-Activating Enzymes/metabolism Deubiquitinating Enzymes/metabolism Endopeptidases Endopeptidases/genetics Animals Humans Protein Precursors Ubiquitins Genetics Multidisciplinary Deubiquitinating Enzymes biology Ubiquitination Protein Precursors/genetics Ubiquitin Thiolesterase/metabolism Protein Precursors/metabolism Ubiquitins/genetics Ubiquitin precursor Protein ubiquitination Ubiquitin ligase Cell biology De novo synthesis Deubiquitinating Enzymes/genetics USP9X Ubiquitin/biosynthesis Liver biology.protein Ribosomal Proteins/metabolism Rabbits Ubiquitin/metabolism Ubiquitin Thiolesterase Protein Processing Post-Translational HeLa Cells |
| Zdroj: | Scientific Reports Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| Popis: | Protein ubiquitination, a major post-translational modification in eukaryotes, requires an adequate pool of free ubiquitin. Cells maintain this pool by two pathways, both involving deubiquitinases (DUBs): recycling of ubiquitin from ubiquitin conjugates and processing of ubiquitin precursors synthesized de novo. Although many advances have been made in recent years regarding ubiquitin recycling, our knowledge on ubiquitin precursor processing is still limited, and questions such as when are these precursors processed and which DUBs are involved remain largely unanswered. Here we provide data suggesting that two of the four mammalian ubiquitin precursors, UBA < inf > 52 < /inf > and UBA < inf > 80 < /inf > , are processed mostly post-translationally whereas the other two, UBB and UBC, probably undergo a combination of co-and post-translational processing. Using an unbiased biochemical approach we found that UCHL < inf > 3 < /inf > , USP < inf > 9 < /inf > X, USP < inf > 7 < /inf > , USP < inf > 5 < /inf > and Otulin/Gumby/FAM < inf > 105 < /inf > b are by far the most active DUBs acting on these precursors. The identification of these DUBs together with their properties suggests that each ubiquitin precursor can be processed in at least two different manners, explaining the robustness of the ubiquitin de novo synthesis pathway. We are grateful to Dr. Cheryl Arrowsmith (University of Toronto, Canada) for providing the plasmids pET28a-LIC-USP5 (Addgene plasmid 25299) and pET28a-LIC-USP5(C335A). We thank Dr. João M. Cabral (IBMC, University of Porto, Portugal) for critically reading the manuscript. This work was supported by national funds through FCT - Fundação para a Ciência e a Tecnologia/MEC – Ministério da Educação e Ciência and when applicable co-funded by Fundo de Desenvolvimento Regional (FEDER) funds within the partnership agreement PT2020 related with the research unit number 4293; by Project “NORTE-07-0124-FEDER-000003 -Cell homeotasis tissue organization and organism biology”, co-funded by Programa Operacional Regional do Norte (ON.2—O Novo Norte), under the Quadro de Referência Estratégico Nacional (QREN), through FEDER and by FCT; by Portuguese National Mass Spectrometry Network (RNEM) through the project REDE/1504/REM/2005; and by Química Orgânica, Produtos Naturais e Agroalimentares (QOPNA) research unit funds provided by FCT, European Union, QREN, FEDER and Operational Competitiveness Programme (COMPETE) under the projects PEst-C/QUI/UI0062/2013 and FCOMP-01-0124-FEDER-037296. C.P.G. and M.P.P. were supported by FCT, COMPETE and Fundo Social Europeu. A.V.M. was supported by the project FCOMP-01-0124-FEDER-027627-EXPL/BEX-BCM/0320/2012 financed by national funds from FCT/Ministério da Educação e Ciência (PIDDAC) and co-funded by FEDER through COMPETE—Programa Operacional Factores de Competitividade (POFC). |
| Databáze: | OpenAIRE |
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