2012.01.12 Cysteine methylation disrupts ubiquitin-chain sensing in NF-kB activation.pdf

LETTER doi:10.1038/nature10690 Cysteine methylation disrupts ubiquitin-chain sensing in NF-kB activation Li Zhang1,2, Xiaojun Ding2, Jixin Cui2, Hao Xu2, Jing Chen2, Yi-Nan Gong2, Liyan Hu2, Yan Zhou2, Jianning Ge2, Qiuhe Lu2, Liping Liu2, She Chen2 Feng Shao2 NF-kB is crucial for innate immune defence against microbial infection1,2. Inhibition of NF-kB signalling has been observed with various bacterial infections3,4. The NF-kB pathway critically requires multiple ubiquitin-chain signals of different natures5,6. Thequestionofwhetherubiquitin-chain signalling and its specificity in NF-kB activation are regulated during infection, and how this regulation takes place, has not been explored. Here we show that human TAB2 and TAB3, ubiquitin-chain sensory proteins involved in NF-kB signalling, are directly inactivated by enteropathogenic Escherichia coli NleE, a conserved bacterial type-III-secreted effector responsible for blocking host NF-kB signalling. NleE harboured an unprecedented S-adenosyl-L-methionine-dependent methyltrans- ferase activity that specifically modified a zinc-coordinating cysteine in the Npl4 zinc finger (NZF) domains in TAB2 and TAB3. Cysteine-methylated TAB2-NZF andTAB3-NZF (truncated proteins only comprising the NZF domain) lost the zinc ion as well as the ubiquitin-chain binding activity. Ectopically expressed or type-III-secretion-system-delivered NleE methylated TAB2 and TAB3 in host cells and diminished their ubiquitin-chain binding activity. Replacement of the NZF domain of TAB3 with the NleE methylation-insensitiveNpl4NZFdomain resulted inNleE-resistant NF-kB activation. Given the prevalence of zinc-finger motifs and activation of cysteine thiol by zinc binding, methylation of zinc- finger cysteinemight regulate other eukaryotic pathways in addition to NF-kB signalling. Bacterial pathogens such as Salmonella,Shigella andenteropathogenic E. coli (EPEC) target the NF-kB pathway to counteract host defence3,4,7. Recent genetic studies reveal