Cloning and characterization of a homologue of the alpha inhibitor of NF-kappaB in Rainbow trout (Oncorhynchus mykiss).

A homologue of IkappaBalpha, the alpha member of the IkappaB family of NF-kappaB inhibitors, was identified in a Rainbow trout suppression subtractive hybridization library enriched in sequences up-regulated in cultured leukocytes after lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNFalpha) stimulation. The full-length cDNA was isolated and sequenced. The predicted amino acid sequence is 61.5% similar and 54% identical to human IkappaBalpha, while only 42% similar and 35% identical to IkappaBbeta, and 38% similar and 32% identical to IkappaBvarepsilon. Rainbow trout IkappaBalpha contains a central ankyrin repeat domain required for its interaction with NF-kappaB and a putative PEST-like sequence in the C-terminus. Expression of IkappaBalpha is up-regulated by LPS and TNFalpha treatment, two known activators of NF-kappaB, suggesting the existence of an autoregulatory loop in fish, as is the case for mammals. These results confirm the existence of the NF-kappaB signalling pathway in fish and suggest a similar functional interaction between IkappaBalpha and NF-kappaB.

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