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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.

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