Espinosa Joaquín 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
Facultad de Ciencias Exactas y Naturales, Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Buenos Aires, Argentina.
Mol Biochem Parasitol. 2003 Sep;131(1):35-44. 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)(2)Cys(X)(4)His(X)(4)Cys. A recombinant PZFP1 was generated in E. coli and the expected 21kDa polypeptide co-purified with two other inducible products of about 42 and 63kDa. Western blot analysis of cell extracts using an anti-PZFP1 antibody recognized a major band of 41kDa. 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.
首次在克氏锥虫中鉴定出一种名为PZFP1的多锌指蛋白。该蛋白有191个氨基酸，包含七个Cys(X)(2)Cys(X)(4)His(X)(4)Cys基序。在大肠杆菌中产生了重组PZFP1，预期的21kDa多肽与另外两种约42kDa和63kDa的可诱导产物共纯化。使用抗PZFP1抗体对细胞提取物进行的蛋白质印迹分析识别出一条41kDa的主要条带。电泳迁移率变动分析表明，重组PZFP1和天然PZFP1均与携带其他CCHC蛋白识别序列的单链DNA或RNA寡核苷酸特异性相互作用。通过间接免疫荧光分析观察到该蛋白主要定位于细胞质和细胞核中。在酵母双杂交试验中，PZFP1与克氏锥虫富含丝氨酸-精氨酸的蛋白（TcSR）特异性相互作用，表明其在mRNA前体加工中发挥作用。
