Genetic construction and characterization of the diphtheria toxin-related interleukin 15 fusion protein DAB389 sIL-15.
A gene fusion encoding DAB389 sIL-15 was constructed in which the catalytic and transmembrane domains of native diphtheria toxin (DAB389) are genetically linked to the N-terminus of simian interleukin 15 (sIL-15). It was demonstrated that the cytotoxic action of DAB389 sIL-15 is mediated through the IL-15 receptor. Since toxicity may be blocked with chloroquine, it was concluded that following binding to the IL-15 receptor, the fusion toxin is internalized by receptor-mediated endocytosis and must pass through an acidic compartment in order to facilitate the delivery of the catalytic domain to the cytosol of target cells. As a non-toxic control, the ADP-ribosyltransferase defective mutant DA(E149S)B389 sIL-15 was constructed. It was demonstrated that both sIL-15 and DA(E149S)B389 sIL-15 stimulate protein and DNA synthesis in IL-15 receptor-positive CTLL-2 cells in vitro.
vanderSpek
J C
JC
Evans Department of Clinical Research, Boston University Medical Center Hospital, MA 02118, USA.
Sutherland
J
J
Sampson
E
E
Murphy
J R
JR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004167:Diphtheria Toxin / Q000737:chemistry* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D000882:Haplorhini / Q000378:metabolism; D019409:Interleukin-15 / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D011485:Protein Binding; D015202:Protein Engineering; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D018123:Receptors, Interleukin; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D017421:Sequence Analysis; D015398:Signal Transduction
D004167:Diphtheria Toxin; D019409:Interleukin-15; D011506:Proteins; D018123:Receptors, Interleukin; D011993:Recombinant Fusion Proteins; D004247:DNA