Recently, potent toxic plant lectins have been conjugated to monoclonal antibodies directed against specific cell surface determinants to provide new avenues for selective cancer therapy. The manner in which these compounds exert their lethal effects in the disruption of protein synthesis by the ribosome is unknown. New techniques to compre reibosomal structural components form toxin resistant cells to toxin-sensitive cells have the potential to answer this question. We will generate yeast strains with toxin-resistant ribosomes by chemical mutagen tratement or by the introduction of genes from toxin-resistant plants. Resistnat strains will be selected by their ability to be transformed by a genetic construct which expresses the toxin, grown in the presence of toxin, and ultimately, maintain protein synthesis after direct ribosome treatment by toxin. Protein chromatographic and RNA sequencing techniques will then be used to define the basis for resistance, providing insight into the mechanisms of protein synthesis. This course of study not only addresses a fundamental problem shared by the disciplines of biochemistry and medicine, but it also incorporates a wide variety of essential scientific techniques vital for the development of independent research skills. As such, the refinement of the skills and observation encompassed in this inquiry will serve as a complement to formal graduate study in biochemistry and molecular biology. In turn, this foundation of basic training and expertise will underlie the future initiation and execution of independent research endeavors which apply this specific understanding of cellular function and molecular biology to clinical problems in medicine.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Physician Scientist Award (K11)
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Allergy & Clinical Immunology-1 (AITC)
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University of Minnesota Twin Cities
Schools of Medicine
United States
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Perentesis, J P; Waddick, K G; Bendel, A E et al. (1997) Induction of apoptosis in multidrug-resistant and radiation-resistant acute myeloid leukemia cells by a recombinant fusion toxin directed against the human granulocyte macrophage colony-stimulating factor receptor. Clin Cancer Res 3:347-55
Perentesis, J P; Bendel, A E; Shao, Y et al. (1997) Granulocyte-macrophage colony-stimulating factor receptor-targeted therapy of chemotherapy- and radiation-resistant human myeloid leukemias. Leuk Lymphoma 25:247-56
Perentesis, J P; Gunther, R; Waurzyniak, B et al. (1997) In vivo biotherapy of HL-60 myeloid leukemia with a genetically engineered recombinant fusion toxin directed against the human granulocyte macrophage colony-stimulating factor receptor. Clin Cancer Res 3:2217-27
Bendel, A E; Shao, Y; Davies, S M et al. (1997) A recombinant fusion toxin targeted to the granulocyte-macrophage colony-stimulating factor receptor. Leuk Lymphoma 25:257-70
Phan, L D; Perentesis, J P; Bodley, J W (1993) Saccharomyces cerevisiae elongation factor 2. Mutagenesis of the histidine precursor of diphthamide yields a functional protein that is resistant to diphtheria toxin. J Biol Chem 268:8665-8
Perentesis, J P; Phan, L D; Gleason, W B et al. (1992) Saccharomyces cerevisiae elongation factor 2. Genetic cloning, characterization of expression, and G-domain modeling. J Biol Chem 267:1190-7
Perentesis, J P; Miller, S P; Bodley, J W (1992) Protein toxin inhibitors of protein synthesis. Biofactors 3:173-84
Perentesis, J P; Genbauffe, F S; Veldman, S A et al. (1988) Expression of diphtheria toxin fragment A and hormone-toxin fusion proteins in toxin-resistant yeast mutants. Proc Natl Acad Sci U S A 85:8386-90