Silkworm coatomers and their role in tube expansion of posterior silkgland.

Silkworm coatomers and their role in tube expansion of posterior silkgland.
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蚕外壳及其在后丝腺管扩张中的作用。

DOI:
10.1371/journal.pone.0013252
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发表时间:
2010-10-12
期刊:
影响因子:
3.7
通讯作者:
Chen J
Chen J
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Wang Q;Shen B;Zheng P;Feng H;Chen L;Zhang J;Zhang C;Zhang G;Teng J;Chen J

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外壳蛋白复合物I(COPI)囊泡,由七个外壳体亚基包被,主要负责高尔基体到内质网的运输。家蚕后丝腺(PSG)是一种高度分化的分泌组织,分泌丝素蛋白用于生产蚕丝,但PSG细胞中的许多生理过程有待进一步研究。为了研究家蚕COPI的作用,我们克隆了6个家蚕COPI亚基(α、β、β′、δ、ε和β-COP),测定了它们在5龄PSG中2天的表达峰值,并观察了COPI作为一种外套复合物与顺式高尔基体的定位。通过将dsRNA注射到家蚕幼虫体内,我们抑制了α-、β′-和γ-COP的表达,证明COPI亚基是PSG管扩张所必需的。α-COP的敲除破坏了高尔基体的完整性,导致PSG的腺腔变窄,表明家蚕COPI是PSG管扩张所必需的。初步表征揭示了家蚕COPI在PSG中的重要作用。虽然蚕COPI类似于先前在其他生物体中表征的被衣分子,但一些令人惊讶的发现需要进一步研究。因此,我们的研究结果表明,家蚕作为研究细胞内运输的模型,并将有助于建立家蚕PSG作为一个有效的生物反应器。
Coat protein complex I (COPI) vesicles, coated by seven coatomer subunits, are mainly responsible for Golgi-to-ER transport. Silkworm posterior silkgland (PSG), a highly differentiated secretory tissue, secretes fibroin for silk production, but many physiological processes in the PSG cells await further investigation. Here, to investigate the role of silkworm COPI, we cloned six silkworm COPI subunits (α,β,β′, δ, ε, and ζ-COP), determined their peak expression in day 2 in fifth-instar PSG, and visualized the localization of COPI, as a coat complex, with cis-Golgi. By dsRNA injection into silkworm larvae, we suppressed the expression of α-, β′- and γ-COP, and demonstrated that COPI subunits were required for PSG tube expansion. Knockdown of α-COP disrupted the integrity of Golgi apparatus and led to a narrower glandular lumen of the PSG, suggesting that silkworm COPI is essential for PSG tube expansion. The initial characterization reveals the essential roles of silkworm COPI in PSG. Although silkworm COPI resembles the previously characterized coatomers in other organisms, some surprising findings require further investigation. Therefore, our results suggest the silkworm as a model for studying intracellular transport, and would facilitate the establishment of silkworm PSG as an efficient bioreactor.
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