title: Expression profile and subcellular localization of HslV, the proteasome related protease from Trypanosoma cruzi.
abstract: Trypanosoma cruzi is a rare example of an eukaryote that has genes for two threonine proteases: HslVU complex and 20S proteasome. HslVU is an ATP-dependent protease consisting of two multimeric components: the HslU ATPase and the HslV peptidase. In this study, we expressed and obtained specific antibodies to HslU and HslV recombinant proteins and demonstrated the interaction between HslU/HslV by coimmunoprecipitation. To evaluate the intracellular distribution of HslV in T. cruzi we used an immunofluorescence assay and ultrastructural localization by transmission electron microscopy. Both techniques demonstrated that HslV was localized in the kinetoplast of epimastigotes. We also analyzed the HslV/20S proteasome co-expression in Y, Berenice 62 (Be-62) and Berenice 78 (Be-78) T. cruzi strains. Our results showed that HslV and 20S proteasome are differently expressed in these strains. To investigate whether a proteasome inhibitor could modulate HslV and proteasome expressions, epimastigotes from T. cruzi were grown in the presence of PSI, a classical proteasome inhibitor. This result showed that while the level of expression of HslV/20S proteasome is not affected in Be-78 strain, in Y and Be-62 strains the presence of PSI induced a significantly increase in Hslv/20S proteasome expression. Together, these results suggest the coexistence of the protease HslVU and 20S proteasome in T. cruzi, reinforcing the hypothesis that non-lysosomal degradation pathways have an important role in T. cruzi biology.
authors.affiliation: Departamento de Ciências Biológicas & NUPEB, Instituto de Ciências Exatas e Biológica, Universidade Federal de Ouro Preto, Morro do Cruzeiro, 35400-000 Ouro Preto, MG, Brazil. nrbarboza@gmail.com
authors.forename: Natália Rocha
authors.initials: NR
authors.lastname: Barboza
authors.forename: Josiane
authors.initials: J
authors.lastname: Cardoso
authors.forename: Carla V
authors.initials: CV
authors.lastname: de Paula Lima
authors.forename: Maurilio J
authors.initials: MJ
authors.lastname: Soares
authors.forename: Daniela Fiore
authors.initials: DF
authors.lastname: Gradia
authors.forename: Nilza Satie
authors.initials: NS
authors.lastname: Hangai
authors.forename: Maria T
authors.initials: MT
authors.lastname: Bahia
authors.forename: Marta
authors.initials: M
authors.lastname: de Lana
authors.forename: Marco A
authors.initials: MA
authors.lastname: Krieger
authors.forename: Renata
authors.initials: R
authors.lastname: Guerra de Sá
mesh_terms: D049069:ATP-Dependent Proteases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D015971:Gene Expression Regulation, Enzymologic; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D051379:Mice; D008928:Mitochondria / Q000201:enzymology; D009842:Oligopeptides / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D061988:Proteasome Inhibitors; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure
chemical_list: D009842:Oligopeptides; D061988:Proteasome Inhibitors; C090535:benzyloxycarbonyl-isoleucyl-glutamyl(O-tert-butyl)-alanyl-leucinal; D049069:ATP-Dependent Proteases; D046988:Proteasome Endopeptidase Complex