title: New insights into host-parasite ubiquitin proteome dynamics in P. falciparum infected red blood cells using a TUBEs-MS approach.
abstract: Malaria, caused by Plasmodium falciparum (P. falciparum), ranks as one of the most baleful infectious diseases worldwide. New antimalarial treatments are needed to face existing or emerging drug resistant strains. Protein degradation appears to play a significant role during the asexual intraerythrocytic developmental cycle (IDC) of P. falciparum. Inhibition of the ubiquitin proteasome system (UPS), a major intracellular proteolytic pathway, effectively reduces infection and parasite replication. P. falciparum and erythrocyte UPS coexist during IDC but the nature of their relationship is largely unknown. We used an approach based on Tandem Ubiquitin-Binding Entities (TUBEs) and 1D gel electrophoresis followed by mass spectrometry to identify major components of the TUBEs-associated ubiquitin proteome of both host and parasite during ring, trophozoite and schizont stages. Ring-exported protein (REX1), a P. falciparum protein located in Maurer's clefts and important for parasite nutrient import, was found to reach a maximum level of ubiquitylation in trophozoites stage. The Homo sapiens (H. sapiens) TUBEs associated ubiquitin proteome decreased during the infection, whereas the equivalent P. falciparum TUBEs-associated ubiquitin proteome counterpart increased. Major cellular processes such as DNA repair, replication, stress response, vesicular transport and catabolic events appear to be regulated by ubiquitylation along the IDC P. falciparum infection.
  
  
  CONCLUSIONS
  In this work we analyze for the first time the interconnection between Plasmodium and human red blood cells ubiquitin-regulated proteins in the context of infection. We identified a number of human and Plasmodium proteins whose ubiquitylation pattern changes during the asexual infective stage. We demonstrate that ubiquitylation of REX1, a P. falciparum protein located in Maurer's clefts and important for parasite nutrient import, peaks in trophozoites stage. The ubiquitin-proteome from P. falciparum infected red blood cells (iRBCs) revealed a significant host-parasite crosstalk, underlining the importance of ubiquitin-regulated proteolytic activities during the intraerythrocytic developmental cycle (IDC) of P. falciparum. Major cellular processes defined from gene ontology such as DNA repair, replication, stress response, vesicular transport and catabolic events appear to be regulated by ubiquitylation along the IDC P. falciparum infection. Given the importance of ubiquitylation in the development of infectious diseases, this work provides a number of potential drug-target candidates that should be further explored.
authors.affiliation: Medicines Development Campus, Diseases of the Developing World, GlaxoSmithKline Tres Cantos, Madrid, Spain; Proteomics Platform CICbioGUNE, CIBERehd, ProteoRed-ISCIII, Parque Tecnologico de Bizkaia, Derio, Spain; Ubiquitylation and Cancer Molecular Biology, Inbiomed, San Sebastian, Spain.
authors.forename: Lydia
authors.initials: L
authors.lastname: Mata-Cantero
authors.affiliation: Proteomics Platform CICbioGUNE, CIBERehd, ProteoRed-ISCIII, Parque Tecnologico de Bizkaia, Derio, Spain.
authors.forename: Mikel
authors.initials: M
authors.lastname: Azkargorta
authors.affiliation: Ubiquitylation and Cancer Molecular Biology, Inbiomed, San Sebastian, Spain.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Aillet
authors.affiliation: Proteomics Platform CICbioGUNE, CIBERehd, ProteoRed-ISCIII, Parque Tecnologico de Bizkaia, Derio, Spain.
authors.forename: Wendy
authors.initials: W
authors.lastname: Xolalpa
authors.affiliation: Medicines Development Campus, Diseases of the Developing World, GlaxoSmithKline Tres Cantos, Madrid, Spain.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: LaFuente
authors.affiliation: Proteomics Platform CICbioGUNE, CIBERehd, ProteoRed-ISCIII, Parque Tecnologico de Bizkaia, Derio, Spain.
authors.forename: Felix
authors.initials: F
authors.lastname: Elortza
authors.affiliation: Computational and Experimental Biology Group, Health Promotion and Chronic Diseases Department, National Institute of Health Dr Ricardo Jorge, Lisbon, Portugal.
authors.forename: Ana Sofia
authors.initials: AS
authors.lastname: Carvalho
authors.affiliation: Centro de Investigación Básica, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
authors.forename: Julio
authors.initials: J
authors.lastname: Martin-Plaza
authors.affiliation: Computational and Experimental Biology Group, Health Promotion and Chronic Diseases Department, National Institute of Health Dr Ricardo Jorge, Lisbon, Portugal. Electronic address: runem2009@gmail.com.
authors.forename: Rune
authors.initials: R
authors.lastname: Matthiesen
authors.affiliation: Proteomics Platform CICbioGUNE, CIBERehd, ProteoRed-ISCIII, Parque Tecnologico de Bizkaia, Derio, Spain; Ubiquitylation and Cancer Molecular Biology, Inbiomed, San Sebastian, Spain; Institut des Technologies Avancées en sciences du Vivant (ITAV), Université de Toulouse, CNRS, UPS, France; University of Toulouse III-Paul Sabatier, 31077 Toulouse, France; Institut de Pharmacologie et de Biologie Structurale (IPBS), Université de Toulouse, CNRS, UPS, France. Electronic address: manuel.rodriguez@itav.fr.
authors.forename: Manuel S
authors.initials: MS
authors.lastname: Rodriguez
mesh_terms: D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology; D006790:Host-Parasite Interactions / Q000502:physiology*; D006801:Humans; D016778:Malaria, Falciparum / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000502:physiology*; D020543:Proteome / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D020543:Proteome; D015800:Protozoan Proteins; D025801:Ubiquitin
keywords: Drug targets; Malaria; Mass spectrometry; Plasmodium falciparum; TUBEs; Ubiquitin