The ubiquitin-like modifier FAT10 decorates autophagy-targeted Salmonella and contributes to Salmonella resistance in mice

V. Spinnenhirn, H. Farhan, M. Basler, A. Aichem, A. Canaan, M. Groettrup
2014 Journal of Cell Science  
Bacterial invasion of eukaryotic cells is counteracted by cellautonomous innate immune mechanisms including xenophagy. The decoration of cytosolic bacteria by ubiquitylation and binding of galectin-8 leads to recruitment of autophagy adaptors like p62 (also known as SQSTM1), NDP52 (also known as CALCOCO2) and optineurin, which initiate the destruction of bacteria by xenophagy. Here, we show that the functionally barely characterized IFNc-and TNFa-inducible ubiquitin-like modifier FAT10 (also
more » ... wn as ubiquitin D, UBD), which binds to the autophagy adaptor p62, but has not been shown to associate with pathogens before, is recruited to cytosolic Salmonella Typhimurium in human cells. FAT10-decorated S. Typhimurium were simultaneously decorated with ubiquitin, p62, NDP52 and the autophagy marker LC3B (MAP1LC3B). FAT10 colocalized with p62-positive microdomains on S. Typhimurium, whereas colocalization with NDP52 was only partial. A kinetic analysis revealed an early, but only transient, decoration of bacteria by FAT10, which resembled that of p62. Although bacterial replication was not detectably altered in FAT10-depleted or overexpressing cells in vitro, survival experiments revealed that NRAMP1-transgenic mice that were FAT10-deficient had a higher susceptibility to orally inoculated S. Typhimurium bacteria than NRAMP1-transgenic mice that were wild-type for FAT10. Taken together, our data suggest a role for FAT10 in the intracellular defense against bacteria. Cells were grown on glass coverslips and stimulated with 200 U/ml IFNc and 400 U/ml TNFa for 24 h to induce FAT10 expression. Cells were fixed for 15 min with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100 for 10 min at room temperature and blocked with 0.2%
doi:10.1242/jcs.152371 pmid:25271057 fatcat:oqhjfzbngbccnkmtp5ei6gmj5u