title: ADP-ribosylation of translation elongation factor 2 by diphtheria toxin in yeast inhibits translation and cell separation.
abstract: Eukaryotic translation elongation factor 2 (eEF2) facilitates the movement of the peptidyl tRNA-mRNA complex from the A site of the ribosome to the P site during protein synthesis. ADP-ribosylation (ADP(R)) of eEF2 by bacterial toxins on a unique diphthamide residue inhibits its translocation activity, but the mechanism is unclear. We have employed a hormone-inducible diphtheria toxin (DT) expression system in Saccharomyces cerevisiae which allows for the rapid induction of ADP(R)-eEF2 to examine the effects of DT in vivo. ADP(R) of eEF2 resulted in a decrease in total protein synthesis consistent with a defect in translation elongation. Association of eEF2 with polyribosomes, however, was unchanged upon expression of DT. Upon prolonged exposure to DT, cells with an abnormal morphology and increased DNA content accumulated. This observation was specific to DT expression and was not observed when translation elongation was inhibited by other methods. Examination of these cells by electron microscopy indicated a defect in cell separation following mitosis. These results suggest that expression of proteins late in the cell cycle is particularly sensitive to inhibition by ADP(R)-eEF2.
authors.affiliation: Department of Biochemistry and Molecular Biology, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA.
authors.forename: Maria K
authors.initials: MK
authors.lastname: Mateyak
authors.forename: Terri Goss
authors.initials: TG
authors.lastname: Kinzy
mesh_terms: D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D004167:Diphtheria Toxin / Q000096:biosynthesis* / Q000235:genetics; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D011132:Polyribosomes / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004271:DNA, Fungal; D004167:Diphtheria Toxin; D015852:Eukaryotic Initiation Factor-2; D029701:Saccharomyces cerevisiae Proteins; D000244:Adenosine Diphosphate
keywords: ADP-ribosylation; Bacterial Toxins; Cell Cycle; Diphthamide; Protein Synthesis; Translation Elongation Factors