Identification of PDZ5, a candidate universal minicircle sequence binding protein of Trypanosoma cruzi.
The dodecamer universal minicircle sequence is a conserved sequence present in minicircles of trypanosomatid kinetoplast DNA studied so far. This sequence is recognised by a protein named universal minicircle sequence binding protein, described for Crithidia fasciculata, involved in minicircle DNA replication. We have identified a Trypanosoma cruzi gene homologue of the Crithidia fasciculata universal minicircle sequence binding protein. Similar to the Crithidia fasciculata universal minicircle sequence binding protein, the Trypanosoma cruzi protein, named PDZ5, contains five zinc finger motifs. Pulsed field gel electrophoresis indicated that the pdz5 gene is located in the chromosomal band XX of the Trypanosoma cruzi genome. The predicted amino acid sequence of PDZ5 shows a high degree of similarity with several trypanosomatid zinc finger proteins. Specific antibody raised against Crithidia fasciculata universal minicircle sequence binding protein recognises both the recombinant and endogenous PDZ5. The complete pdz5 coding sequence cloned in bacteria expresses a recombinant PDZ5 protein that binds specifically to the universal minicircle sequence dodecamer. These data strongly suggest that PDZ5 represents a Trypanosoma cruzi universal minicircle sequence binding protein.
Coelho
Elielton R
ER
Programa de Biologia Molecular, Instituto de Biofísica Carlos Chagas Filho, CCS, bloco G Universidade Federal de Rio de Janeiro, 21949-900 Rio de Janeiro, Brazil.
Urményi
Turan P
TP
Franco da Silveira
José
J
Rondinelli
Edson
E
Silva
Rosane
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D018105:DNA, Kinetoplast / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000235:genetics*; D016335:Zinc Fingers
D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C095676:universal minicircle sequence binding protein, protozoan