The zinc finger protein TcZFP2 binds target mRNAs enriched during Trypanosoma cruzi metacyclogenesis

• Published in: Memórias do Instituto Oswaldo Cruz 2012-09, Vol.107 (6), p.790-799
• Main Authors: 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
• Format: Article
• Language: English
• Subjects: CCCH zinc finger protein; cell differentiation; DNA-Binding Proteins - metabolism; Gene Expression Regulation, Developmental; PARASITOLOGY; Protozoan Proteins - metabolism; RNA Stability; RNA, Messenger - metabolism; RNA-binding protein; RNA-Binding Proteins - metabolism; TROPICAL MEDICINE; Trypanosoma cruzi; Trypanosoma cruzi - CCCH zinc finger protein - RNA-binding protein - cell differentiation; Trypanosoma cruzi - growth & development; Trypanosoma cruzi - metabolism
• ISSN: 1678-8060; 0074-0276; 1678-8060; 0074-0276
• DOI: 10.1590/S0074-02762012000600014

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 . 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 protein levels showed ZFP2 to be a positive regulator of specific target mRNA abundance.