UDP-glucose deficiency causes hypersensitivity to the cytotoxic effect of Clostridium perfringens phospholipase C

被引:30
作者
Flores-Díaz, M
Alape-Girón, A
Titball, RW
Moos, M
Guillouard, I
Cole, S
Howells, AM
von Eichel-Streiber, C
Florin, I
Thelestam, M
机构
[1] Karolinska Inst, Microbiol & Tumorbiol Ctr, S-17177 Stockholm, Sweden
[2] Univ Costa Rica, Fac Med, Dept Bioquim, San Jose, Costa Rica
[3] Chem & Biol Def Estab, Def Evaluat & Res Agcy, Salisbury SP4 0JQ, Wilts, England
[4] Inst Med Mikrobiol & Hyg, D-55101 Mainz, Germany
[5] Inst Pasteur, Unite Genet Mol Bacterienne, F-75724 Paris 15, France
关键词
D O I
10.1074/jbc.273.38.24433
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 10(5) times more sensitive to the cytotoxic effect of Clostridium perfringens phospholipase 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.
引用
收藏
页码:24433 / 24438
页数:6
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