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Factors from Trypanosoma cruzi interacting with AP-1 sequences
J Espinosa1, H Martinetto, D Portal
1Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad de Buenos Aires, Argentina.
The Journal of Eukaryotic Microbiology
|October 16, 1999
Summary
This study investigated Trypanosoma cruzi factors binding to AP-1 DNA sequences using electrophoretic mobility shift assays. AP-1 binding activity was detected in all parasite forms, with higher levels in amastigote and trypomastigote stages, and was enhanced by oxidative stress.
Area of Science:
- Molecular biology
- Parasitology
- Biochemistry
Background:
- The protozoan parasite Trypanosoma cruzi causes Chagas disease.
- Understanding host-parasite interactions at the molecular level is crucial for therapeutic development.
- Transcription factor AP-1 plays a role in various cellular processes, including parasite development.
Purpose of the Study:
- To investigate the interaction between Trypanosoma cruzi factors and AP-1 DNA binding sequences.
- To characterize the nature and abundance of AP-1 binding activity in different parasite forms.
Main Methods:
- Electrophoretic mobility shift assays (EMSA) were employed using double- and single-stranded DNA probes containing the AP-1 consensus sequence.
- Complexes were stabilized by UV cross-linking and analyzed by SDS-PAGE.
- Antibodies against mammalian c-Jun and c-Fos were used for protein identification.
Main Results:
- Three specific complexes (A, B, C) were detected, with complex B specific for double-stranded DNA.
- AP-1 binding activity was higher in nuclear preparations and present in all three parasite forms (epimastigote, amastigote, trypomastigote).
- Binding was significantly stronger in amastigote and trypomastigote forms and increased under oxidative stress. Proteins of 40- and 60-kDa, potentially homologous to c-Jun and c-Fos, were identified.
Conclusions:
- Trypanosoma cruzi possesses AP-1 binding factors that interact with specific DNA sequences.
- The abundance and activity of these factors vary across parasite life cycle stages and are influenced by environmental conditions like oxidative stress.
- These findings suggest a potential role for AP-1-like transcription factors in the biology and pathogenesis of Trypanosoma cruzi.