UDP-glucose Deficiency Causes Hypersensitivity to the Cytotoxic Effect of Clostridium perfringens Phospholipase C

• Published in: The Journal of biological chemistry Vol. 273; no. 38; pp. 24433 - 24438
• Main Authors: Flores-Dı́az, Marietta, Alape-Girón, Alberto, Titball, Richard W., Moos, Michael, Guillouard, Isabelle, Cole, Stewart, Howells, Angela M., von Eichel-Streiber, Christoph, Florin, Inger, Thelestam, Monica
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.09.1998; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Antibodies, Monoclonal - pharmacology; Bacterial Toxins - biosynthesis; Bacterial Toxins - chemistry; Bacterial Toxins - toxicity; Calcium-Binding Proteins; Cattle; Cell Line; Cell Membrane - drug effects; Cell Membrane - physiology; Cell Survival - drug effects; Cell Survival - physiology; Choline - metabolism; Clostridium - enzymology; Clostridium perfringens; Clostridium perfringens - enzymology; Cricetinae; Cricetulus; Kinetics; Lung; Medicin och hälsovetenskap; Membrane Lipids - biosynthesis; Peptide Fragments - pharmacology; Recombinant Proteins - biosynthesis; Recombinant Proteins - metabolism; Recombinant Proteins - toxicity; Transfection; Type C Phospholipases - biosynthesis; Type C Phospholipases - chemistry; Type C Phospholipases - toxicity; Uridine Diphosphate Glucose - deficiency; Uridine Diphosphate Glucose - metabolism; UTP-Glucose-1-Phosphate Uridylyltransferase - deficiency; UTP-Glucose-1-Phosphate Uridylyltransferase - genetics; UTP-Glucose-1-Phosphate Uridylyltransferase - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.38.24433

A Chinese hamster cell line with a mutation in the UDP-glucose pyrophosphorylase (UDPG:PP) gene leading to UDP-glucose deficiency as well as a revertant cell were previously isolated. We now show that the mutant cell is 105 times more sensitive to the cytotoxic effect of Clostridium perfringensphospholipase C (PLC) than the revertant cell. To clarify whether there is a connection between the UDP-glucose deficiency and the hypersensitivity to C. perfringens PLC, stable transfectant cells were prepared using a wild type UDPG:PP cDNA. Clones of the mutant transfected with a construct having the insert in the sense orientation had increased their UDP-glucose level, whereas those of the revertant transfected with a UDPG:PP antisense had reduced their level of UDP-glucose compared with control clones transfected with the vector. Exposure of these two types of transfectant clones to C. perfringens PLC demonstrated that a cellular UDP-glucose deficiency causes hypersensitivity to the cytotoxic effect of this phospholipase. Further experiments with genetically engineered C. perfringens PLC variants showed that the sphingomyelinase activity and the C-domain are required for its cytotoxic effect in UDP-glucose-deficient cells.