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Identification and function analysis of BCL2 in immune response of Pteria penguin
Wenhao He1, Jinji Lin1, Feifei Yu2
1College of Fisheries, Guangdong Ocean University, Zhanjiang, Guangdong, 524088, China; Academician Joint Laboratory of Germplasm Resource Exploitation, Utilization and Health Assessment for Aquatic Animal, Guangdong Ocean University, Zhanjiang, Guangdong, 524088, China.
Abstract:
B-cell lymphoma/leukemia-2 (BCL2), an anti-apoptotic factor in the mitochondrial regulatory pathway of apoptosis, is critically important in immune defenses. In this study, a novel BCL2 gene was characterized from Pteria penguin (P. penguin). The PpBCL2 was 1482 bp long, containing an open reading frame (ORF) of 588 bp encoding 195 amino acids. Four highly conserved BCL-2 homology (BH) domains were found in PpBCL2. Amino acid alignment and phylogenetic tree showed that PpBCL2 had the highest similarity with BCL2 of Crassostrea gigas at 65.24 %. Tissue expression analysis showed that PpBCL2 had high constitutive expression in gill, digestive diverticulum and mantle, and was significantly increased 72 h of Vibrio parahaemolyticus (V. parahaemolyticus) challenge in these immune tissues. Furthermore, PpBCL2 silencing significantly inhibited antimicrobial activity of hemolymph supernatant by 1.4-fold, and significantly reduced the survival rate by 51.7 % at 72 h post infection in P. penguin. These data indicated that PpBCL2 played an important role in immune response of P. penguin against V. parahaemolyticus infection.
Insights
This study identified a novel BCL2 gene in Pteria penguin, crucial for immune defense. Its expression increased during Vibrio parahaemolyticus infection, highlighting its role in combating bacterial challenges.
Area of Science:
- Marine Biology
- Immunology
- Molecular Biology
Background:
- The B-cell lymphoma/leukemia-2 (BCL2) gene is a key anti-apoptotic factor regulating immune responses.
- Understanding BCL2's role in invertebrates is vital for immune defense mechanisms.
Purpose of the Study:
- To characterize a novel BCL2 gene from Pteria penguin (PpBCL2).
- To investigate the function of PpBCL2 in P. penguin's immune response against Vibrio parahaemolyticus.
Main Methods:
- Gene cloning and sequencing of Pteria penguin BCL2 (PpBCL2).
- Bioinformatic analysis including amino acid alignment and phylogenetic tree construction.
- Gene silencing via RNA interference and assessment of antimicrobial activity and survival rates post-infection.
Main Results:
- A novel PpBCL2 gene of 1482 bp with an ORF of 588 bp encoding 195 amino acids was identified, featuring four conserved BCL-2 homology domains.
- PpBCL2 showed highest similarity (65.24%) to BCL2 from Crassostrea gigas.
- PpBCL2 was constitutively expressed in immune tissues (gill, digestive diverticulum, mantle) and significantly upregulated upon Vibrio parahaemolyticus challenge.
- Silencing PpBCL2 reduced hemolymph antimicrobial activity by 1.4-fold and decreased survival rates by 51.7% after 72 hours of infection.
Conclusions:
- PpBCL2 plays a significant role in the immune defense of Pteria penguin against Vibrio parahaemolyticus infection.
- The study provides insights into the molecular mechanisms of invertebrate immunity involving BCL2 family proteins.
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