The T300A Crohn's disease risk polymorphism impairs function of the WD40 domain of ATG16L1
A coding polymorphism of human ATG16L1 (rs2241880; T300A) increases the risk of Crohn's disease and it has been shown to enhance susceptibility of ATG16L1 to caspase cleavage. Here we show that T300A also alters the ability of the C-terminal WD40-repeat domain of ATG16L1 to interact with an amino acid motif that recognizes this region. Such alteration impairs the unconventional autophagic activity of TMEM59, a transmembrane protein that contains the WD40 domain-binding motif, and disrupts its normal intracellular trafficking and its ability to engage ATG16L1 in response to bacterial infection. TMEM59-induced autophagy is blunted in cells expressing the fragments generated by caspase processing of the ATG16L1-T300A risk allele, whereas canonical autophagy remains unaffected. These results suggest that the T300A polymorphism alters the function of motif-containing molecules that engage ATG16L1 through the WD40 domain, either by influencing this interaction under non-stressful conditions or by inhibiting their downstream autophagic signalling after caspase-mediated cleavage. ; This work was funded by grants from the Ministerio de Ciencia e Innovación (Ref. SAF2011-23714) and the Ministerio de Economía y Competitividad (Ref. SAF2014-53320-R) of the Spanish Government, the Broad Medical Research Program at Crohn's and Colitis Foundation of America (CCFA; Ref. IBD-0369), the Junta de Castilla y León local government (Department of Education (Refs. CSI001A10-2, FIC016U14) and Department of Health (Ref. SAN11-FXP)) and the Fundación Solorzano (Refs. FS/1-2009 and FS/18-2014). R.J.X. is funded by the NIH (grants R01DK097485, P30DK043351 and U19AI109725) and the Helmsley Trust. Additional funding comes from the FEDER programme of the European Union. E.B.R. and I.S.G. are graduate students funded by predoctoral fellowships from the FPU programme (Ministerio de Educación, MEC, Spanish Government) and the Fundación Moraza, respectively. ; Peer Reviewed