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

Memórias do Instituto Oswaldo Cruz

Patricia Alves Mörking²  Rita De Cássia Pontello Rampazzo²  Pegine Walrad¹  Christian Macagnan Probst²  Maurilio José Soares²  Daniela Fiori Gradia²  Daniela Parada Pavoni²  Marco Aurélio Krieger²  Keith Matthews¹  Samuel Goldenberg²  Stenio Perdigão Fragoso²  Bruno Dallagiovanna² 

[1] ,Fiocruz Instituto Carlos Chagas Curitiba PR ,Brasil

关键词: Trypanosoma cruzi;    CCCH zinc finger protein;    RNA-binding protein;    cell differentiation;   

DOI  :  10.1590/S0074-02762012000600014

来源: SciELO

【 摘 要 】

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 culture-derived 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.

【 授权许可】

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