Trypanosoma cruzi poly-zinc finger protein: a novel DNA/RNA-binding CCHC-zinc finger protein

• Published in: Molecular and biochemical parasitology 2003-09, Vol.131 (1), p.35-44
• Main Authors: Espinosa, Joaquı́n M, Portal, Daniel, Lobo, Guillermo S, Pereira, Claudio A, Alonso, Guillermo D, Gómez, Eliana B, Lan, Graciela Humbert, Pomar, Rolando V.Rivera, Flawiá, Mirtha M, Torres, Héctor N
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; CCHC-type zinc finger; Cloning, Molecular; DNA, Protozoan - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Humans; Molecular Sequence Data; Poly-zinc finger protein; Protozoan Proteins - chemistry; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; PZFP1 protein; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Protozoan - metabolism; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Sequence Alignment; Serine-arginine-rich protein; TcPZFP1; TcSR; Trypanosoma cruzi; Trypanosoma cruzi - chemistry; Trypanosoma cruzi - genetics; Trypanosoma cruzi - metabolism; Two-Hybrid System Techniques; Zinc Fingers
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/S0166-6851(03)00187-7

A poly-zinc finger protein, designated PZFP1 was identified in Trypanosoma cruzi for the first time. The protein has 191 amino acids, contains seven motifs Cys(X) 2Cys(X) 4His(X) 4Cys. A recombinant PZFP1 was generated in E. coli and the expected 21 kDa polypeptide co-purified with two other inducible products of about 42 and 63 kDa. Western blot analysis of cell extracts using an anti-PZFP1 antibody recognized a major band of 41 kDa. Electrophoretic mobility shift analysis demonstrated that both, recombinant and native PZFP1, specifically interact with single-stranded DNA or RNA oligonucleotides carrying recognition sequences of other CCHC proteins. The protein was localized mainly in the cytoplasm and nucleus as observed by indirect immunofluorescence analysis. PZFP1 interacted specifically with a T. cruzi serine–arginine-rich protein (TcSR) in a yeast two-hybrid assay, suggesting a role in pre-mRNA processing.