ATG16L1的编码等位基因会增加克罗恩病的风险(T300A;rs2241880)损害C端WD40结构域(WDD)与含有WDD结合基序的蛋白质之间的相互作用,从而特异性抑制ATG16L1的非常规自噬活动。在最近的出版物中,我们描述了ATG16L1在调节IL10R(白介素10受体)运输和信号传导中的新型非典型作用。涉及WDD与IL10RB(白介素10受体亚基β)中存在的靶基序之间直接相互作用的活性。在这里我们展示,出乎意料的是,T300A突变不会改变ATG16L1与IL10RB相互作用的能力及其在支持IL10信号传导中的作用。这些结果表明,ATG16L1T300A等位基因选择性地削弱了WDD与WDD结合基序版本的子集之间的相互作用,这表明只有一小部分由ATG16L1介导的非常规活性是预防克罗恩病所必需的。缩写:ATG,自噬相关;ATG16L1,自噬相关16样1;BMDMs,骨髓源性巨噬细胞;CRISPR,成簇的规则间隔的短回文重复序列;CSF1/M-CSF,集落刺激因子1;FBS,胎牛血清;GSH,谷胱甘肽;IL10,白细胞介素10;IL10R,白细胞介素10受体;LPS,脂多糖;MAP1LC3/LC3,微管相关蛋白1轻链3;MEFs,小鼠胚胎成纤维细胞;PMA,肉豆蔻酸佛波醇乙酸酯;p-STAT3:磷酸化STAT3;qPCR,定量聚合酶链反应;SDS,十二烷基硫酸钠;sgRNA,单向导RNA;TMEM59,跨膜蛋白59;TNF,肿瘤坏死因子;TNFAIP3/A20,TNFα诱导蛋白3;WDD,WD40结构域;WIPI2,WD重复结构域,磷酸肌醇相互作用2.
A coding allele of ATG16L1 that increases the risk of Crohn disease (T300A; rs2241880) impairs the interaction between the C-terminal WD40 domain (WDD) and proteins containing a WDD-binding motif, thus specifically inhibiting the unconventional autophagic activities of ATG16L1. In a recent publication we described a novel atypical role of ATG16L1 in the regulation of IL10R (interleukin 10 receptor) trafficking and signaling, an activity that involves direct interaction between the WDD and a target motif present in IL10RB (interleukin 10 receptor subunit beta). Here we show that, unexpectedly, neither the ability of ATG16L1 to interact with IL10RB nor its role in supporting IL10 signaling are altered by the T300A mutation. These results indicate that the ATG16L1T300A allele selectively impairs the interaction between the WDD and a subset of WDD-binding motif versions, suggesting that only a fraction of the unconventional activities mediated by ATG16L1 are required to prevent Crohn disease.Abbreviations: ATG, autophagy related; ATG16L1, autophagy related 16 like 1; BMDMs, bone marrow-derived macrophages; CRISPR, clustered regularly interspaced short palindromic repeats; CSF1/M-CSF, colony stimulating factor 1; FBS, fetal bovine serum; GSH, glutathione; IL10, interleukin 10; IL10R, interleukin 10 receptor; LPS, lipopolysaccharide; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; MEFs, mouse embryonic fibroblasts; PMA, phorbol myristate acetate; p-STAT3: phosphorylated STAT3; qPCR, quantitative polymerase chain reaction; SDS, sodium dodecyl sulfate; sgRNA, single guide RNA; TMEM59, transmembrane protein 59; TNF, tumor necrosis factor; TNFAIP3/A20, TNF alpha induced protein 3; WDD, WD40 domain; WIPI2, WD repeat domain, phosphoinositide interacting 2.