Functional complementation of glycoprotein 72 in a Trypanosoma cruzi glycoprotein 72 null mutant.
To investigate the role of a developmentally regulated 72-kDa surface antigen of Trypanosoma cruzi (GP72), a GP72 null mutant was previously produced [Cooper et al., 1993, J. Cell Biol. 122, 149-156]. Abnormal morphology of epimastigote and metacyclic trypomastigote stages of the GP72 null mutant suggested that GP72 is associated with flagellum-cell adhesion [Cooper et al., 1993, J. Cell Biol. 122, 149-156; De Jesus et al., J. Cell Sci., in press]. In the present study, functional complementation of the GP72 null mutant was accomplished by transformation with two independent episomal vectors expressing GP72 and phleomycin or tunicamycin resistance genes. A correlation between gene copy number, RNA level, expression of GP72, and morphological phenotypes was demonstrated. Disparities were observed between gene copy number and RNA level and between the apparent level of GP72 polypeptide and the carbohydrate epitope recognized by monoclonal antibody WIC29.26. Restoration of morphology reflects recovery of the carbohydrate epitope, suggesting that the novel glycosylation of GP72 is the limiting step in the expression of its function.
Laboratory of Molecular Parasitology, Rockefeller University, New York, NY 10021.
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D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000378:metabolism; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics; D005784:Gene Amplification; D017125:Genes, Protozoan; D005816:Genetic Complementation Test; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D010692:Phleomycins / Q000494:pharmacology; D010750:Phosphoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000254:growth & development / Q000276:immunology; D014415:Tunicamycin / Q000494:pharmacology
D000953:Antigens, Protozoan; D000954:Antigens, Surface; C038790:GP72 antigen, Trypanosoma; D010692:Phleomycins; D010750:Phosphoproteins; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D014415:Tunicamycin