Smurf participates in Helicoverpa armigera diapause by regulating the transforming growth factor-β signaling pathway.

Abstract

Diapause, an important strategy used by insects to avoid adverse environments, is regulated by various cell signaling pathways. The results of our previous studies demonstrated that the transforming growth factor-β (TGF-β) signaling pathway regulated pupal diapause in Helicoverpa armigera, which was accompanied by downregulation of proteins in TGF-β signaling. However, to date the mechanism underlying this phenomenon remains unknown. Here, we cloned the E3 ubiquitin ligases gene Smurf. In vitro experiments showed that Smurf directly bound to TGF-β receptor type I (TGFβRI) and Smad2. Overexpressing Smurf promoted ubiquitination of TGFβRI and Smad2, thereby downregulating their protein levels. Conversely, silencing of the Smurf gene suppressed ubiquitination of TGFβRI and Smad2 thereby increasing their protein levels. Results from in vivo co-immunoprecipitation assays revealed that the binding of Smurf to TGFβRI or Smad2 was stronger in diapause pupae than in nondiapause pupae. Injection of Smurf inhibitor A01 into diapause pupae markedly upregulated expression of TGFβRI and Smad2 proteins, leading to resumption of development in diapause pupae. Taken together, these findings suggested that ubiquitin ligase E3 Smurf participated in H. armigera diapause by regulating TGF-β signaling, and thus could be playing a crucial role in insect diapause.