Kinetoplast DNA-binding protein profile in the epimastigote form of Trypanosoma cruzi.
BACKGROUND
The Trypanosomatidae family possesses one of the most unusual DNAs found in nature: the kinetoplast genome. It consists of a few dozen maxicircles that encode for some subunits of mitochondrial enzymes and rRNAs in a cryptic pattern and thousands of minicircles that encode for the guide RNAs (gRNAs), all catenated and constituting a dense network. The complexity of kinetoplast genome based on its intricate DNA structure is well known; however, only a small number of proteins associated with kinetoplast DNA (kDNA) have been described, and the majority are related with the replication process.


METHODS
We describe the protein profile obtained using formaldehyde as a cross-linking agent to obtain the kinetoplast DNA-protein complex, and Southwestern assay to identify the kDNA binding proteins present in the complex.


RESULTS
We identified seven proteins eluted from the kDNA complex fixed by formaldehyde. Polyclonal antiserum developed against the kDNA-protein complex recognized only four proteins in crude extracts of epimastigote stage, suggesting immunogenic differences among these proteins and/or their availability in the kinetoplast genome. Southwestern assay using minicircle fragments showed nine kDNA binding proteins in crude extracts of Trypanosoma cruzi epimastigote.


CONCLUSIONS
We describe several proteins associated with the kDNA. Some could be involved in the essential process for parasite life and also could be a good target for drug or vaccine development. The results contribute to understanding the organization of the kinetoplast genome.
Zavala-Castro
Jorge E
JE
Departamento de Patología Experimental, Centro de Investigaciones de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV del IPN), Mexico City, Mexico. zcastro@tunku.uady.mx
Acosta-Viana
Karla
K
Baylon-Pacheco
Lydia
L
González-Robles
Arturo
A
Guzmán-Marín
Eugenia
E
Rosales-Encina
José L
JL
D000818:Animals; D018105:DNA, Kinetoplast / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015151:Immunoblotting; D014349:Trypanosoma cruzi / Q000378:metabolism*
D018105:DNA, Kinetoplast; D004268:DNA-Binding Proteins