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Published on: April 21, 2015
Altering of host larval (Spodoptera exigua) calcineurin activity in response to ascovirus infection
Huan Yu1,2, Lei He1,2, Zi-Qi Li1,2
1Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha, P.R. China.
Background:
Calcineurin (CaN) is involved in numerous cellular processes and Ca2+ -dependent signal transduction pathways. According to our previous transcriptome studies, thousands of host larval (Spodoptera exigua) transcripts were downregulated after the infection of Heliothis virescent ascovirus 3h (HvAV-3h), while the Spodoptera exigua calcineurin genes (SeCaNs) were significantly upregulated. To understand the regulation of SeCaNs in S. exigua larvae during the infection of HvAV-3h, the functions of CaN subunit A (SeCaN-SubA) and CaN binding protein (SeCaN-BP) were analysed.
Results:
The in vitro assays indicated that the bacterial expressed SeCaN-SubA is an acid phosphatase, but no phosphatase activity was detected with the purified SeCaN-BP. The transcription level of SeCaN-SubA was upregulated after HvAV-3h infection and the CaN activity was significantly increased after HvAV-3h infection in S. exigua larvae. Interestingly, the SeCaN-BP transcripts were only detectable in the HvAV-3h infected larvae. Further immunoblotting results consistently agree with those obtained by qPCR, indicating that the infection of HvAV-3h causes the upregulated expression of SeCaN-SubA and the appearance of SeCaN-BP. An interaction between the cleaved SeCaN-SubA and SeCaN-BP was detected by co-immunoprecipitation assays, and the expression of SeCaN-BP in Spodoptera frugiperda-9 (Sf9) cells can help to increase the CaN activity of SeCaN-SubA. Further investigations with CaN inhibitors suggested that HvAV-3h. Further investigations with CaN inhibitors suggested that the inhibition on host larval CaN activity can also inhibit the viral replication of HvAV-3h.
Conclusion:
The increase in CaN activity caused by HvAV-3h infection might be due to the upregulation of SeCaN-SubA and the induced expression of SeCaN-BP, and increased CaN activity is essential for ascoviral replication. © 2019 Society of Chemical Industry.
Insights
Heliothis virescent ascovirus 3h (HvAV-3h) infection upregulates Spodoptera exigua calcineurin subunit A (SeCaN-SubA) and induces Spodoptera exigua calcineurin binding protein (SeCaN-BP). Increased calcineurin activity is essential for ascoviral replication.
Area of Science:
- Molecular Biology
- Virology
- Insect Pathology
Background:
- Calcineurin (CaN) is a key enzyme in Ca2+-dependent signaling pathways.
- Heliothis virescent ascovirus 3h (HvAV-3h) infection leads to downregulation of host transcripts but upregulation of Spodoptera exigua calcineurin genes (SeCaNs).
Purpose of the Study:
- To investigate the functions of SeCaN subunit A (SeCaN-SubA) and SeCaN binding protein (SeCaN-BP) in S. exigua larvae during HvAV-3h infection.
- To understand the regulation of SeCaNs in response to HvAV-3h infection.
Main Methods:
- In vitro expression and activity assays of SeCaN-SubA and SeCaN-BP.
- Quantitative PCR (qPCR) and immunoblotting to analyze transcript and protein levels.
- Co-immunoprecipitation assays to detect protein interactions.
- Calcineurin inhibitor assays to assess the impact on viral replication.
Main Results:
- Bacterially expressed SeCaN-SubA exhibited acid phosphatase activity, while SeCaN-BP did not.
- HvAV-3h infection upregulated SeCaN-SubA transcription and increased overall CaN activity in larvae.
- SeCaN-BP transcripts were specifically detected in HvAV-3h infected larvae, with its expression increasing CaN activity.
- Co-immunoprecipitation confirmed an interaction between cleaved SeCaN-SubA and SeCaN-BP.
- Inhibition of host larval CaN activity suppressed HvAV-3h replication.
Conclusions:
- HvAV-3h infection increases CaN activity through SeCaN-SubA upregulation and SeCaN-BP induction.
- Elevated CaN activity is crucial for the replication of HvAV-3h.
- Targeting host calcineurin presents a potential strategy for controlling ascoviral infections.
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