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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 107, No. 6, 2012, pp. 790-799
Bioline Code: oc12123
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 107, No. 6, 2012, pp. 790-799

 en The zinc finger protein TcZFP2 binds target mRNAs enriched during Trypanosoma cruzi check for this species in other resources metacyclogenesis
Mörking, Patricia Alves; Rampazzo, Rita de Cássia Pontello; Walrad, Pegine; Probst, Christian Macagnan; Soares, Maurilio José; Gradia, Daniela Fiori; Pavoni, Daniela Parada; Krieger, Marco Aurélio; Matthews, Keith; Goldenberg, Samuel; Fragoso, Stenio Perdigão & Dallagiovanna, Bruno

Abstract

Trypanosomes are parasitic protozoa in which gene expression is primarily controlled through the regulation of mRNA stability and translation. This post-transcriptional control is mediated by various families of RNA-binding proteins, including those with zinc finger CCCH motifs. CCCH zinc finger proteins have been shown to be essential to differentiation events in trypanosomatid parasites. Here, we functionally characterise TcZFP2 as a predicted post-transcriptional regulator of differentiation in Trypanosoma cruzi check for this species in other resources . This protein was detected in cell culturederived amastigotes and trypomastigotes, but it was present in smaller amounts in metacyclic trypomastigote forms of T. cruzi. We use an optimised recombinant RNA immunopreciptation followed by microarray analysis assay to identify TcZFP2 target mRNAs. We further demonstrate that TcZFP2 binds an A-rich sequence in which the adenosine residue repeats are essential for high-affinity recognition. An analysis of the expression profiles of the genes encoding the TcZFP2-associated mRNAs throughout the parasite life cycle by microarray hybridisation showed that most of the associated mRNAs were upregulated in the metacyclic trypomastigote forms, also suggesting a role for TcZFP2 in metacyclic trypomastigote differentiation. Knockdown of the orthologous Trypanosoma brucei check for this species in other resources protein levels showed ZFP2 to be a positive regulator of specific target mRNA abundance.

Keywords
Trypanosoma cruzi - CCCH zinc finger protein - RNA-binding protein - cell differentiation

 
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