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SpBOK inhibits WSSV infection by regulating the apoptotic pathway in mud crab (Scylla paramamosain)
Shanmeng Lin1, Yuyong He1, Yi Gong1
1Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou, 515063, China; Marine Biology Institute, Shantou University, Shantou, 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.
Abstract:
B-cell lymphoma 2 (Bcl-2) related ovarian killer (BOK) is a member of the Bcl-2 family, which has a similar function to BAX and BAK in the process of apoptosis. However, how BOK activates the intrinsic (mitochondrial) apoptotic pathway remains poorly understood in invertebrates. In this study, SpBOK identified in mud crab is an important effector responsible for the anti-WSSV (White Spot Syndrome Virus) infection by activating the apoptotic pathway. The SpBOK gene encoded a 282 amino acid peptides (molecular mass of 29 kD), which contained four distinct Bcl-2 family homology (BH) domains. SpBOK was widely expressed in all tested tissues and up-regulated after WSSV infection in vivo. The role of SpBOK on the anti-WSSV response in mud crab was investigated by using the RNAi approach in vivo. SpBOK exerted a regulatory role in changing the mitochondrial membrane potential (⊿ψm) and activating the caspase signaling and thus induced apoptosis. Moreover, the results showed that WSSV replication in mud crab could be effectively inhibited by SpBOK. Therefore, the results of this study demonstrated that SpBOK can inhibit WSSV infection by regulating the intrinsic apoptosis pathway in mud crab.
Insights
Mud crabs fight White Spot Syndrome Virus (WSSV) using SpBOK, a protein that triggers apoptosis. This discovery reveals a new mechanism for antiviral defense in invertebrates by regulating the mitochondrial apoptotic pathway.
Area of Science:
- Molecular Biology
- Immunology
- Crustacean Research
Background:
- Bcl-2 related ovarian killer (BOK) proteins are involved in apoptosis.
- The role of BOK in invertebrate apoptosis, particularly during viral infections, is not well understood.
- White Spot Syndrome Virus (WSSV) is a significant pathogen affecting crustaceans.
Purpose of the Study:
- To investigate the function of SpBOK in mud crabs against WSSV infection.
- To elucidate the mechanism by which SpBOK activates the intrinsic apoptotic pathway.
- To determine if SpBOK can inhibit WSSV replication.
Main Methods:
- Identification and characterization of the SpBOK gene in mud crabs.
- Analysis of SpBOK expression patterns after WSSV infection.
- RNA interference (RNAi) to study SpBOK function in vivo.
- Assessment of mitochondrial membrane potential and caspase activation.
- Quantification of WSSV replication.
Main Results:
- SpBOK, a 282-amino acid protein with four BH domains, was identified and widely expressed in mud crabs.
- SpBOK expression was upregulated following WSSV infection.
- RNAi-mediated knockdown of SpBOK impaired the induction of apoptosis and increased WSSV replication.
- SpBOK regulates mitochondrial membrane potential and activates caspase signaling, leading to apoptosis.
- SpBOK effectively inhibited WSSV replication in mud crabs.
Conclusions:
- SpBOK is a key effector in the anti-WSSV immune response in mud crabs.
- SpBOK inhibits WSSV infection by activating the intrinsic mitochondrial apoptotic pathway.
- This study provides insights into invertebrate antiviral defense mechanisms involving apoptosis.
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