Detection and localization of small nuclear ribonucleoproteins (snRNPs) in trypanosomatids using anti-m3G antibodies.
This work reports for the first time the identification and immunolocalization, by confocal and conventional indirect immunofluorescence, of m3G epitopes present in ribonucleoproteins of the following trypanosomatids: Trypanosoma cruzi epimastigotes of three different strains, Blastocrithidia ssp., and Leishmania major promastigotes. The identity of these epitopes and hence the specificity of the anti-m3G monoclonal antibody were ascertained through competition reaction with 7-methylguanosine that blocks the Ig binding sites, abolishing the fluorescence in all the parasites tested and showing a specific perinuclear localization of the snRNPs, which suggests their nuclear reimport in the parasites. Using an immunoprecipitation technique, it was also possible to confirm the presence of the trimethylguanosine epitopes in trypanosomatids.
Bosetto
Maira Cegatti
MC
Departamento de Ciencias Biologicas, Faculdade de Ciencias Farmaceuticas, Universidade Estadual Paulista, Sao Paulo, Brasil.
Mortara
Renato Arruda
RA
Ambrósio
Daniela Luz
DL
Passerini
Gabriela Duó
GD
da Silva
Marco Túlio Alves
MT
Okuda
Erica Sayuri
ES
Cicarelli
Regina Maria Barretto
RM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000939:Epitopes / Q000302:isolation & purification*; D019084:Fluorescent Antibody Technique, Indirect; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D017411:Ribonucleoproteins, Small Nuclear / Q000276:immunology / Q000302:isolation & purification*; D014351:Trypanosomatina / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D017411:Ribonucleoproteins, Small Nuclear