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核转运因子2和Ran在棉铃虫烟青虫20E信号转导途径中的功能

Function of nuclear transport factor 2 and Ran in the 20E signal transduction pathway in the cotton bollworm, Helicoverpa armigera.

作者信息

He Hong-Juan, Wang Qian, Zheng Wei-Wei, Wang Jin-Xing, Song Qi-Sheng, Zhao Xiao-Fan

机构信息

School of Life Sciences, Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education, Shandong University, Jinan 250100, Shandong, PR China.

出版信息

BMC Cell Biol. 2010 Jan 2;11:1. doi: 10.1186/1471-2121-11-1.

DOI:10.1186/1471-2121-11-1
PMID:20044931
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2830935/
Abstract

BACKGROUND

Nuclear transport factor 2 and small GTPase Ran participate in the nucleo-cytoplasm transport of macromolecules, but their function in the 20-hydroxyecdysone (20E) signal transduction pathway are not well known.

RESULTS

A 703 bp encoding Ntf2 and a 1233 bp encoding Ran full-length cDNAs were cloned from Helicoverpa armigera, and named Ha-Ntf2 and Ha-Ran, respectively. Northern blot and immunoblotting revealed that Ha-Ntf2 had an obviously higher expression levels in the head-thorax and integument of the metamorphically committed larvae. In contrast, the expression of Ha-Ran did not show obvious variation at various developmental stages in four tissues by immunoblotting analysis, except in the midgut, which showed increased expression from 5th-36 h (molting) to 6th-48 h. Both expressions of Ha-Ntf2 and Ha-Ran could be upregulated by 20E in vitro. Immunohistochemistry revealed that Ha-Ntf2 and Ha-Ran were primarily localized in the nucleus of various tissues. Protein binding assay and co-immunoprecipitation indicated that Ha-Ntf2 and Ha-Ran can combine with each other in vitro and in vivo. Knock down of Ha-Ntf2 or Ha-Ran by RNAi resulted in the suppression of other 20E regulated genes including EcR-B1, USP1, E75B, BR-CZ2, HHR3 and Ha-eIF5c. In addition, the knockdown of Ha-Ntf2 resulted in Ha-Ran being prevented in the cytoplasm. The nuclear location of the ecdysone receptor b1 (EcR-B1) was also blocked after the knockdown of Ha-Ntf2 and Ha-Ran.

CONCLUSION

These evidences suggested that Ha-Ntf2 and Ha-Ran participated in the 20E signal transduction pathway by regulating the location of EcR-B1.

摘要

背景

核转运因子2和小GTP酶Ran参与大分子的核质运输，但其在20-羟基蜕皮酮（20E）信号转导途径中的功能尚不清楚。

结果

从棉铃虫中克隆出编码Ntf2的703 bp和编码Ran的1233 bp全长cDNA，分别命名为Ha-Ntf2和Ha-Ran。Northern杂交和免疫印迹显示，Ha-Ntf2在变态期幼虫的头胸部和体表有明显较高的表达水平。相比之下，通过免疫印迹分析，Ha-Ran在四个组织的不同发育阶段表达没有明显变化，除了中肠，从第5天-36小时（蜕皮）到第6天-48小时表达增加。Ha-Ntf2和Ha-Ran的表达在体外均可被20E上调。免疫组织化学显示，Ha-Ntf2和Ha-Ran主要定位于各组织的细胞核中。蛋白质结合试验和免疫共沉淀表明，Ha-Ntf2和Ha-Ran在体外和体内均可相互结合。通过RNA干扰敲低Ha-Ntf2或Ha-Ran会导致其他20E调控基因（包括EcR-B1、USP1、E75B、BR-CZ2、HHR3和Ha-eIF5c）的表达受到抑制。此外，敲低Ha-Ntf2会导致Ha-Ran滞留在细胞质中。敲低Ha-Ntf2和Ha-Ran后，蜕皮激素受体b1（EcR-B1）的核定位也被阻断。

结论

这些证据表明，Ha-Ntf2和Ha-Ran通过调节EcR-B1的定位参与20E信号转导途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/364ec479d802/1471-2121-11-1-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/70c9c1801678/1471-2121-11-1-1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/2d55b5670077/1471-2121-11-1-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/1d584f4c3b28/1471-2121-11-1-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/88927e8bd61e/1471-2121-11-1-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/267801f27693/1471-2121-11-1-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/05e062e49fb1/1471-2121-11-1-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/364ec479d802/1471-2121-11-1-9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/70c9c1801678/1471-2121-11-1-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/5b80f4f74469/1471-2121-11-1-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/a8905e4903ef/1471-2121-11-1-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/2d55b5670077/1471-2121-11-1-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/1d584f4c3b28/1471-2121-11-1-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/88927e8bd61e/1471-2121-11-1-6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/267801f27693/1471-2121-11-1-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/05e062e49fb1/1471-2121-11-1-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d722/2830935/364ec479d802/1471-2121-11-1-9.jpg

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