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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.

journal_name

Exp Parasitol

journal_title

Experimental parasitology

authors

Barboza NR,Cardoso J,de Paula Lima CV,Soares MJ,Gradia DF,Hangai NS,Bahia MT,de Lana M,Krieger MA,Guerra de Sá R

doi

10.1016/j.exppara.2011.10.011

subject

Has Abstract

pub_date

2012-02-01 00:00:00

pages

171-7

issue

2

eissn

0014-4894

issn

1090-2449

pii

S0014-4894(11)00323-7

journal_volume

130

pub_type

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