Toxin resistance and reduced secretion in a mouse L-cell mutant defective in herpes virus propagation.
The mouse L-cell mutant gro29 was selected originally for its inability to propagate herpes simplex virus; it shows severe defects in virus egress and the transport and processing of viral glycoproteins after infection. In this report, we show that uninfected gro29 cells display pleiotropic changes in protein secretion, oligosaccharide processing, and sensitivity to the toxins ricin and modeccin. Specifically, the rate of secretion of a nonglycosylated protein, human growth hormone, was reduced 70% in gro29 cells compared with the parental L cells. A direct measurement of the transport capacity of Golgi membranes in a cell-free assay suggests that gro29 cells contain less functional Golgi than parental cells. Despite this deficiency, N-linked oligosaccharides were processed efficiently in mutant cells, although there were differences in the structure of the mature forms. Lectin intoxication assays revealed that gro29 cells were cross-resistant to killing by the cytotoxic lectins ricin and modeccin, but not to wheat germ agglutinin, Ricinus communis agglutinin RCA120, or leucoagglutinin. Fluorescence labeling using fluorescein-conjugated lectins showed that uninfected gro29 cells expressed relatively few ricin-binding molecules, suggesting a possible mechanism for toxin resistance. These studies provide evidence that the processes of protein secretion, lectin intoxication, and herpes virus maturation and egress may share a common cellular component.
Michaelis
C
C
Department of Microbiology, University of British Columbia, Vancouver, Canada.
Banfield
B W
BW
Gruenheid
S
S
Tsang
Y
Y
Lippe
R
R
Jefferies
W A
WA
Wattenberg
B W
BW
Tufaro
F
F
D000818:Animals; D001692:Biological Transport / Q000235:genetics; D016466:CHO Cells; D002474:Cell-Free System; D006224:Cricetinae; D004351:Drug Resistance; D006023:Glycoproteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D007739:L Cells / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D037102:Lectins / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018139:Simplexvirus / Q000502:physiology*; D014118:Toxins, Biological / Q000494:pharmacology*; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology
D006023:Glycoproteins; D037102:Lectins; D011993:Recombinant Fusion Proteins; D014118:Toxins, Biological; D014764:Viral Proteins