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TcZFP8, a novel member of the Trypanosoma cruzi CCHC zinc finger protein family with nuclear localization
A.O. Ericsson, L.O. Faria, W.B. Cruz, C. Martins de Sá and B.D. Lima
Laboratório de Biologia do Gene, Departamento de Biologia Celular,
Universidade de Brasília, Brasília, DF, Brasil
Corresponding author: B.D. Lima
E-mail: bdlima@unb.br
Genet. Mol. Res. 5 (3): 553-563 (2006)
Received January 24, 2006
Accepted August 10, 2006
Published September 14, 2006

ABSTRACT. In a 17-kb genomic fragment of Trypanosoma cruzi chromosome XX, we identified three tandemly linked genes coding for CX2CX4HX4C zinc finger proteins. We also showed that similar genes are present in T. brucei and Leishmania major, sharing three monophyletic groups among these trypanosomatids. In T. cruzi, TcZFP8 corresponds to a novel gene coding for a protein containing eight zinc finger motifs. Molecular cloning of this gene and heterologous expression as a fusion with a His-tag were performed in Escherichia coli. The purified recombinant protein was used to produce antibody in rabbits. Using Western blot analysis, we observed the presence of this protein in all three forms of the parasite: amastigote, trypomastigote and epimastigote. An analysis of cytoplasmic and nuclear cell extracts showed that this protein is present in nuclear extracts, and indirect immunofluorescence microscopy confirmed the nuclear localization of TcZFP8. Homologues of TcZFP8 in T. brucei are apparently absent, while one candidate in L. major was identified.

Key words: CCHC-type zinc finger, Nuclear protein, TcZFP8, Trypanosoma cruzi, Trypanosomatid

 

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