The C-Mannosylome of Human Induced Pluripotent Stem Cells Implies a Role for ADAMTS16 C-Mannosylation in Eye Development.
The C-Mannosylome of Human Induced Pluripotent Stem Cells Implies a Role for ADAMTS16 C-Mannosylation in Eye Development.
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DOI：
10.1016/j.mcpro.2021.100092
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Buettner FFR
中科院分区：
文献类型：
--
作者：
Cirksena K;Hütte HJ;Shcherbakova A;Thumberger T;Sakson R;Weiss S;Jensen LR;Friedrich A;Todt D;Kuss AW;Ruppert T;Wittbrodt J;Bakker H;Buettner FFR
C-甘露糖化是色氨酸残基的单一甘露糖修饰，可以影响蛋白质的折叠、分泌和/或功能。到目前为止，只有几种蛋白质被证明是C-甘露糖化的，而且全球评估蛋白质C-甘露糖化的研究很少。为了研究人类诱导多能干细胞的C-甘露糖体，我们利用基于MS的定量蛋白质组学方法比较了C-甘露糖基转移酶DPY19L1或DPY19L3缺失的CRISPR-Cas9突变体与WT人诱导的多能干细胞的分泌组。在这些突变体中，许多蛋白质的分泌减少，包括A去整合素和带有血栓反应蛋白基序的金属蛋白酶16(ADAMTS16)的分泌，这是一种细胞外蛋白酶，以前被报道对斑马鱼眼睛发育中的视裂融合是必不可少的。为了检验这一观察结果的功能相关性，我们利用CRISPR-Cas9将dpy19l1或dpy19l3定位于日本稻鱼Medaka(Oryzias Latipe)的胚胎中。我们观察到，靶向dpy1913部分导致了视裂融合的缺陷，称为缺陷区。我们进一步在细胞模型中表明，DPY19L1和DPY19L3介导了重组表达的凝血酶反应蛋白1重复的ADAMTS16的C-甘露糖化，从而支持其分泌。综上所述，我们的研究结果表明，DPY19L3介导的C-甘露糖化通过辅助细胞外蛋白水解酶ADAMTS16的分泌参与眼睛的发育。ADAMTS16的TSR1可以是C-甘露糖基。HPSCDPY19L1或DPY19L3缺失导致ADAMTS16分泌减少。在青竹中靶向dpy1913有时会导致缺陷症。我们确定ADAMTS16是C-甘露糖化的靶蛋白，并表明这种修饰是ADAMTS16正常分泌所必需的。CRISPR-Cas9针对青冈鱼胚胎中的一个独特的C-甘露糖基转移酶导致眼睛发育缺陷。我们的结论是，这些发育缺陷是由于C-甘露糖缺失时ADAMTS16分泌减少所致。
C-mannosylation is a modification of tryptophan residues with a single mannose and can affect protein folding, secretion, and/or function. To date, only a few proteins have been demonstrated to be C-mannosylated, and studies that globally assess protein C-mannosylation are scarce. To interrogate the C-mannosylome of human induced pluripotent stem cells, we compared the secretomes of CRISPR–Cas9 mutants lacking either the C-mannosyltransferase DPY19L1 or DPY19L3 to WT human induced pluripotent stem cells using MS-based quantitative proteomics. The secretion of numerous proteins was reduced in these mutants, including that of A Disintegrin And Metalloproteinase with ThromboSpondin Motifs 16 (ADAMTS16), an extracellular protease that was previously reported to be essential for optic fissure fusion in zebrafish eye development. To test the functional relevance of this observation, we targeted dpy19l1 or dpy19l3 in embryos of the Japanese rice fish medaka (Oryzias latipes) by CRISPR–Cas9. We observed that targeting of dpy19l3 partially caused defects in optic fissure fusion, called coloboma. We further showed in a cellular model that DPY19L1 and DPY19L3 mediate C-mannosylation of a recombinantly expressed thrombospondin type 1 repeat of ADAMTS16 and thereby support its secretion. Taken together, our findings imply that DPY19L3-mediated C-mannosylation is involved in eye development by assisting secretion of the extracellular protease ADAMTS16. TSR1 of ADAMTS16 can be C-mannosylated. Deletion of DPY19L1 or DPY19L3 in hiPSCs caused reduced secretion of ADAMTS16. Targeting of dpy19l3 in medaka occasionally led to coloboma. We identified ADAMTS16 as a target protein for C-mannosylation and showed that this modification is needed for proper secretion of ADAMTS16. Targeting a distinct C-mannosyltransferase by CRISPR–Cas9 in medaka fish embryos caused defects in eye development. We conclude that these developmental defects are caused by reduced secretion of ADAMTS16 when C-mannosylation is missing.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
DOI：
10.1016/j.bbrc.2018.10.138
发表时间：
2018-12-02
影响因子：
3.1
作者：
Gouyer, Valerie;Demouveaux, Bastien;Desseyn, Jean-Luc
通讯作者：
Desseyn, Jean-Luc
影响因子：
46.9
作者：
Heck, Dirk;Kowalczyk, Monika S.;Yudovich, David;Belizaire, Roger;Puram, Rishi V.;McConkey, Marie E.;Thielke, Anne;Aster, Jon C.;Regev, Aviv;Ebert, Benjamin L.
通讯作者：
Ebert, Benjamin L.
影响因子：
3.5
作者：
Fujiwara, Miho;Kato, Shintaro;Simizu, Siro
通讯作者：
Simizu, Siro
影响因子：
23.9
作者：
Haase, Alexandra;Olmer, Ruth;Martin, Ulrich
通讯作者：
Martin, Ulrich