An understanding of the ways in which microbial toxins and viruses act at the cellular and molecular level will enable better ways of controlling or abrogating their negative effects on cells and tissues to be designed. The purpose of this project is to isolate and study mutant cultured cells, or human and animal origin, in order to investigate mechanisms of Pseudomonas exotoxin A action, replication of animal viruses, receptor-mediated endocytosis, and the cellular pathways and mechanisms involved. Ongoing studies of a group of cell strains already isolated, that are resistant to Pseudomonas toxin and certain viruses because of diverse genetic lesions will be continued. New strains will also be isolated and characterized, using biological, biochemical, immunological, and genetic techniques. Ligands with known or postulated mechanisms of entry, such as well-characterized viruses, lectins, and selected hormones and transport proteins, will be used as probes to determine the biochemical and genetic basis of resistance in the mutant cells. Monoclonal antibodies to the Pseudomonas toxin receptor will be prepared, in order to isolate, purify, and quantify the receptor, to study its structure-function relationships, and to use as probes for cloning the toxin receptor gene. Monoclonal antibodies to Sindbis virus glycoproteins will be prepared for the purpose of localizing, within the cell, the site of an endopeptidase activity essential for the processing of these glycoproteins, and, it is postulated, for the activation of Pseudomonas toxin. The gene (or genes) for this enzyme activity will be cloned, taking advantage of a group of resistance mutant cells that are deficient in this activity. A unique opportunity to investigate not only sensitivity and resistance to microbial toxins, but also regulation of gene expression and cellular function, and viral replication, is presented.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI009100-24
Application #
2059104
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1979-05-01
Project End
1994-04-30
Budget Start
1993-05-01
Budget End
1994-04-30
Support Year
24
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Vermont & St Agric College
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
05405
Ackermann, Mark R (2014) Lamb model of respiratory syncytial virus-associated lung disease: insights to pathogenesis and novel treatments. ILAR J 55:4-15
Grosz, Drew D; van Geelen, Albert; Gallup, Jack M et al. (2014) Sucrose stabilization of Respiratory Syncytial Virus (RSV) during nebulization and experimental infection. BMC Res Notes 7:158
Derscheid, Rachel J; van Geelen, Albert; Berkebile, Abigail R et al. (2014) Increased concentration of iodide in airway secretions is associated with reduced respiratory syncytial virus disease severity. Am J Respir Cell Mol Biol 50:389-97
Derscheid, Rachel J; Gallup, Jack M; Knudson, Cory J et al. (2013) Effects of formalin-inactivated respiratory syncytial virus (FI-RSV) in the perinatal lamb model of RSV. PLoS One 8:e81472
Inocencio, N M; Moehring, J M; Moehring, T J (1993) A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A is unable to process the precursor fusion glycoprotein of Newcastle disease virus. J Virol 67:593-5
Moehring, J M; Inocencio, N M; Robertson, B J et al. (1993) Expression of mouse furin in a Chinese hamster cell resistant to Pseudomonas exotoxin A and viruses complements the genetic lesion. J Biol Chem 268:2590-4
Robertson, B J; Moehring, J M; Moehring, T J (1993) Defective processing of the insulin receptor in an endoprotease-deficient Chinese hamster cell strain is corrected by expression of mouse furin. J Biol Chem 268:24274-7
Foley, B T; Moehring, J M; Moehring, T J (1992) A mutation in codon 717 of the CHO-K1 elongation factor 2 gene prevents the first step in the biosynthesis of diphthamide. Somat Cell Mol Genet 18:227-31
Fendrick, J L; Iglewski, W J; Moehring, J M et al. (1992) Characterization of the endogenous ADP-ribosylation of wild-type and mutant elongation factor 2 in eukaryotic cells. Eur J Biochem 205:25-31
Watson, D G; Moehring, J M; Moehring, T J (1991) A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A and alphaviruses fails to cleave Sindbis virus glycoprotein PE2. J Virol 65:2332-9

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