Characterization, expression and function analysis of pfTLR5S and pfTLR5M in yellow catfish (Pelteobagrus fulvidraco) responding to bacterial challenge.

Toll-Like Receptors (TLRs) are important pattern recognition receptors, playing critical roles in the early innate immune response to defensing against pathogen invasion. In this study, we found both soluble form TLR5 (pfTLR5S) and membrane form TLR5 (pfTLR5M) in yellow catfish Pelteobagrus fulvidraco. The open reading frames (ORFs) of pfTLR5M and pfTLR5S genes were 2655 bp and 1947 bp in length, encoding 884 and 648 amino acids, respectively. pfTLR5M was composed of thirteen LRR domains, one TIR domain and one transmembrane domain. However, pfTLR5S have only fifteen LRR domains, without any TIR domain and transmembrane domain. Both pfTLR5M and pfTLR5S genes had the highest expression in liver, especially for pfTLR5S, which showed a noticeable high expression in liver. We also compared the relative mRNA expression levels of pfTLR5M and pfTLR5S in digestive and immune-related tissues after challenge of three different bacteria. In addition, we also found that pfTLR5S can interact with pfTLR5M, and inhibit the expression of pfTLR5M protein, while induced the expression of downstream proinflammatory factors, such as TNFα and IL8. These results revealed that both pfTLR5M and pfTLR5S play important and different roles in defensing against the invasion of flagellated bacteria, and they may function by binding to each other.