title: The Hsp90 co-chaperone p23 of Toxoplasma gondii: Identification, functional analysis and dynamic interactome determination.
abstract: Toxoplasma gondii is among the most successful parasites, with nearly half of the human population chronically infected. Recently a link between the T. gondii Hsp90 chaperone machinery and parasite development was observed. Here, the T. gondii Hsp90 co-chaperones p23 and Hip were identified mining the Toxoplasma- database (www.toxodb.org). Their identity was confirmed by domain structure and blast analysis. Additionally, analysis of the secondary structure and studies on the chaperone function of the purified protein verified the p23 identity. Studies of co-immunoprecipitation (co-IP) identified two different types of complexes, one comprising at least Hip-Hsp70-Hsp90 and another containing at least p23-Hsp90. Indirect immunofluorescence assays showed that Hip is localized in the cytoplasm in tachyzoites and as well in bradyzoites. For p23 in contrast, a solely cytoplasmic localization was only observed in the tachyzoite stage whereas nuclear and cytosolic distribution and co-localization with Hsp90 was observed in bradyzoites. These results indicate that the T. gondii Hsp90-heterocomplex cycle is similar to the one proposed for higher eukaryotes, further highlighting the implication of the Hsp90/p23 in parasite development. Furthermore, co-IP experiments of tachyzoite/bradyzoite lysates with anti-p23 antiserum and identification of the complexed proteins together with the use of the curated interaction data available from different source (orthologs and Plasmodium databases) allowed us to construct an interaction network (interactome) covering the dynamics of the Hsp90 chaperone machinery.
authors.affiliation: Laboratorio de Parasitología Molecular, UB2, IIB-INTECH, CONICET-UNSAM, Camino de Circunvalación Laguna Km. 6, C.C 164, (B7130IIWA) Chascomús, Prov. Buenos Aires, Argentina.
authors.forename: Pablo C
authors.initials: PC
authors.lastname: Echeverria
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Figueras
authors.forename: Malvina
authors.initials: M
authors.lastname: Vogler
authors.forename: Thomas
authors.initials: T
authors.lastname: Kriehuber
authors.forename: Natalia
authors.initials: N
authors.lastname: de Miguel
authors.forename: Bin
authors.initials: B
authors.lastname: Deng
authors.forename: Maria C
authors.initials: MC
authors.lastname: Dalmasso
authors.forename: Dwight E
authors.initials: DE
authors.lastname: Matthews
authors.forename: Mariana
authors.initials: M
authors.lastname: Matrajt
authors.forename: Martin
authors.initials: M
authors.lastname: Haslbeck
authors.forename: Johannes
authors.initials: J
authors.lastname: Buchner
authors.forename: Sergio O
authors.initials: SO
authors.lastname: Angel
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry; D019295:Computational Biology; D003593:Cytoplasm / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D018832:Molecular Chaperones / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017433:Protein Structure, Secondary; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014122:Toxoplasma / Q000235:genetics* / Q000378:metabolism*
chemical_list: D016054:DNA, Protozoan; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; C105426:Hip chaperone; D018832:Molecular Chaperones; D015800:Protozoan Proteins
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