title: Protein and mRNA content of TcDHH1-containing mRNPs in Trypanosoma cruzi.
abstract: In trypanosomatids, the regulation of gene expression occurs mainly at the post-transcriptional level. Previous studies have revealed nontranslated mRNA in the Trypanosoma cruzi cytoplasm. Previously, we have identified and cloned the TcDHH1 protein, a DEAD box RNA helicase. It has been reported that Dhh1 is involved in multiple RNA-related processes in various eukaryotes. It has also been reported to accumulate in stress granules and processing bodies of yeast, animal cells, Trypanosoma brucei and T. cruzi. TcDHH1 is localized to discrete cytoplasmic foci that vary depending on the life cycle status and nutritional conditions. To study the composition of mRNPs containing TcDHH1, we carried out immunoprecipitation assays with anti-TcDHH1 using epimastigote lysates. The protein content of mRNPs was determined by MS and pre-immune serum was used as control. We also carried out a ribonomic approach to identify the mRNAs present within the TcDHH1 immunoprecipitated complexes. For this purpose, competitive microarray hybridizations were performed against negative controls, the nonprecipitated fraction. Our results showed that mRNAs associated with TcDHH1 in the epimastigote stage are those mainly expressed in the other forms of the T. cruzi life cycle. These data suggest that mRNPs containing TcDHH1 are involved in mRNA metabolism, regulating the expression of at least epimastigote-specific genes.
authors.affiliation: Instituto Carlos Chagas/FIOCRUZ, Curitiba, Brazil.
authors.forename: Fabíola B
authors.initials: FB
authors.lastname: Holetz
authors.forename: Lysangela R
authors.initials: LR
authors.lastname: Alves
authors.forename: Christian M
authors.initials: CM
authors.lastname: Probst
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallagiovanna
authors.forename: Fabricio K
authors.initials: FK
authors.lastname: Marchini
authors.forename: Patricio
authors.initials: P
authors.lastname: Manque
authors.forename: Gregory
authors.initials: G
authors.lastname: Buck
authors.forename: Marco A
authors.initials: MA
authors.lastname: Krieger
authors.forename: Alejandro
authors.initials: A
authors.lastname: Correa
authors.forename: Samuel
authors.initials: S
authors.lastname: Goldenberg
mesh_terms: D000818:Animals; D015153:Blotting, Western; D053487:DEAD-box RNA Helicases / Q000737:chemistry* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D008018:Life Cycle Stages; D040081:Protein Array Analysis; D012333:RNA, Messenger / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
chemical_list: D012333:RNA, Messenger; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D053487:DEAD-box RNA Helicases