Waterfield Nicholas R, Daborn Phillip J, Dowling Andrea J, Yang Guowei, Hares Michelle, ffrench-Constant Richard H
Department of Biology and Biochemistry, University of Bath BA2 7AY, UK.
FEMS Microbiol Lett. 2003 Dec 12;229(2):265-70. doi: 10.1016/S0378-1097(03)00846-2.
The Photorhabdus luminescens W14 toxin encoding gene makes caterpillars floppy (mcf) was discovered due to its ability to kill caterpillars when expressed in Escherichia coli. Here we describe a homologue of mcf (renamed as mcf1), termed mcf2, discovered in the same genome. The mcf2 gene predicts another large toxin whose central domain, like Mcf1, also shows limited homology to Clostridium cytotoxin B. However, the N-terminus of Mcf2 shows significant similarity to the type-III secreted effector HrmA from the plant pathogen Pseudomonas syringae and no similarity to the N-terminus of Mcf1. HrmA is a plant avirulence gene whose transient expression in tobacco cells results in cell death. Here we show that E. coli expressing Mcf2 can, like E. coli expressing Mcf1, kill insects. Further, expression of the c-Myc tagged N-terminus of Mcf2, the region showing similarity to HrmA, results in nuclear localisation of the fusion protein and subsequent destruction of transfected mammalian cells. The Mcf1 and Mcf2 toxins therefore belong to a family of high molecular mass toxins, differing at their N-termini, which encode different effector domains.
发光杆菌W14的毒素编码基因(使毛虫松弛,mcf)是因其在大肠杆菌中表达时具有杀死毛虫的能力而被发现的。在此,我们描述了在同一基因组中发现的mcf的一个同源物(重新命名为mcf1),称为mcf2。mcf2基因预测了另一种大型毒素,其中心结构域与Mcf1一样,也与梭菌细胞毒素B显示出有限的同源性。然而,Mcf2的N端与植物病原体丁香假单胞菌的III型分泌效应蛋白HrmA具有显著相似性,与Mcf1的N端没有相似性。HrmA是一个植物无毒基因,其在烟草细胞中的瞬时表达会导致细胞死亡。在此我们表明,表达Mcf2的大肠杆菌与表达Mcf1的大肠杆菌一样能够杀死昆虫。此外,与HrmA显示出相似性的Mcf2的c-Myc标记N端的表达会导致融合蛋白的核定位以及随后对转染的哺乳动物细胞的破坏。因此,Mcf1和Mcf2毒素属于一类高分子量毒素家族,它们的N端不同,编码不同的效应结构域。