Entamoeba histolytica: A unicellular organism containing two active genes encoding for members of the TBP family
| Autor: | Carlos Alberto Castañon-Sanchez, Esther Orozco, Maria Esther Herrera-Aguirre, Jose M Hernandez, Ma. Guadalupe de Dios-Bravo, José de Jesús Olivares-Trejo, Juan Pedro Luna-Arias |
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| Rok vydání: | 2010 |
| Předmět: |
Genetics
biology General transcription factor Binding protein Entamoeba histolytica Molecular Sequence Data Protozoan Proteins Electrophoretic Mobility Shift Assay Locus (genetics) TATA-Box Binding Protein biology.organism_classification Genome Molecular biology law.invention law parasitic diseases Recombinant DNA TATA Box Binding Protein-Like Proteins Amino Acid Sequence Cloning Molecular Gene Transcription factor Phylogeny Biotechnology |
| Zdroj: | Protein Expression and Purification. 70:48-59 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2009.12.007 |
| Popis: | Entamoeba histolytica is the protozoan parasite which causes human amoebiasis. In this parasite, few encoding genes for transcription factors have been cloned and characterized. The E. histolytica TATA-box binding protein (EhTBP) is the first basal transcription factor that has been studied. To continue with the identification of other members of the basal transcription machinery, we performed an in silico analysis of the E. histolytica genome and found three loci encoding for polypeptides with similarity to EhTBP. One locus has a 100% identity to the previously Ehtbp gene reported by our group. The second locus encodes for a 212 aa polypeptide that is 100% identical to residues 23-234 from EhTBP. The third one encodes for a 216 aa polypeptide of 24kDa that showed 42.6% identity and 73.7% similarity to EhTBP. This protein was named E. histolytica TBP-related factor 1 (EhTRF1). Ehtrf1 gene was expressed in bacteria and the purified 28kDa recombinant polypeptide showed the capacity to bind to TATTTAAA-box by electrophoretic mobility shift assays. K(D) values for rEhTBP and rEhTRF1 were (1.71+/-2.90)x10(-12)M and (1.12+/-0.160)x10(-11)M, respectively. Homology modeling of EhTRF1 and EhTBP revealed that, although they were very similar, they showed some differences on their surfaces. Thus, E. histolytica is a unicellular organism having two members of the TBP family. |
| Databáze: | OpenAIRE |
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