Expression analysis and functional characterization of the mouse cysteine-rich with EGF-like domains 2.
Expression analysis and functional characterization of the mouse cysteine-rich with EGF-like domains 2.
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DOI：
10.1038/s41598-018-30362-4
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发表时间：
2018-08-15
影响因子：
4.6
通讯作者：
Hirata Y
中科院分区：
文献类型：
--
作者：
Oh-Hashi K;Fujimura K;Norisada J;Hirata Y
我们以前已经发现了一种新的内质网应激诱导蛋白，即富含半胱氨酸的EGF样域2(CRELD2)，它主要受ATF6的调控。然而，关于CRELD2的研究报道很少。在本研究中，我们阐明了内源性CRELD2在小鼠组织和内质网应激处理的Neuro2a细胞中的表达。在我们测试的九种组织中，心脏和骨骼肌中的CRELD2蛋白可以忽略不计。与快速诱导生长停滞和DNA损伤诱导基因153(GADD153)相比，衣霉素(TM)处理Neuro2a细胞后CRELD2的表达在晚期被诱导。另一方面，另一种内质网应激诱导剂thapsigargin增加了Neuro2a细胞内源性CRELD2的分泌。我们进一步建立了CRELD2缺失的Neuro2a细胞来评估它们的特性。与旨在检测活细胞中蛋白质-蛋白质相互作用的纳米Luc互补报告系统相结合，CRELD2不仅与CRELD2本身相互作用，还与Neuro2a细胞中的ER定位蛋白相互作用。最后，我们研究了CRELD2缺陷细胞对TM处理的反应性，发现CRELD2缺陷并不影响由三个典型的ER应激传感器触发的基因的表达，但使Neuro2a细胞对TM刺激敏感。综上所述，这些发现提供了CRELD2的新的分子特征，其进一步的表征将为理解内质网稳态和内质网应激诱导的细胞功能障碍提供新的见解。
We have previously identified a novel endoplasmic reticulum (ER) stress-inducible protein, namely, cysteine-rich with EGF-like domains 2 (CRELD2), which is predominantly regulated by ATF6. However, few studies on intrinsic CRELD2 have been published. In the present study, we elucidated the expression of intrinsic CRELD2 in mouse tissues and ER stress- treated Neuro2a cells. Among nine tissues we tested, CRELD2 protein in the heart and skeletal muscles was negligible. CRELD2 expression in Neuro2a cells was induced at the late phase after treatment with tunicamycin (Tm) compared with rapid induction of growth arrest and DNA damage inducible gene 153 (GADD153). On the other hand, another ER stress inducer, thapsigargin, increased the intrinsic CRELD2 secretion from Neuro2a cells. We furthermore established CRELD2-deficient Neuro2a cells to evaluate their features. In combination with the NanoLuc complementary reporter system, which was designed to detect protein-protein interaction in living cells, CRELD2 interacted with not only CRELD2 itself but also with ER localizing proteins in Neuro2a cells. Finally, we investigated the responsiveness of CRELD2-deficient cells against Tm-treatment and found that CRELD2 deficiency did not affect the expression of genes triggered by three canonical ER stress sensors but rendered Neuro2a cells vulnerable to Tm-stimulation. Taken together, these findings provide the novel molecular features of CRELD2, and its further characterization would give new insights into understanding the ER homeostasis and ER stress-induced cellular dysfunctions.
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影响因子：
3.7
作者：
Norisada J;Hirata Y;Amaya F;Kiuchi K;Oh-hashi K
通讯作者：
Oh-hashi K
影响因子：
19
作者：
Helenius, Ari;Marquardt, Thorsten;Braakman, Ineke
通讯作者：
Braakman, Ineke
影响因子：
21.3
作者：
Murakami, Tomohiko;Saito, Atsushi;Imaizumi, Kazunori
通讯作者：
Imaizumi, Kazunori
影响因子：
64.8
作者：
Calfon, M;Zeng, HQ;Ron, D
通讯作者：
Ron, D
DOI：
10.1126/science.1232033
发表时间：
2013-02-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mali P;Yang L;Esvelt KM;Aach J;Guell M;DiCarlo JE;Norville JE;Church GM
通讯作者：
Church GM