Abstract
14-3-3 family proteins are adaptor proteins that specifically bind to a discrete phosphoserine or phosphothreonine motif in the substrate proteins. They play important roles in metabolism, cell-cycle control, signal transduction, apoptosis, protein trafficking, transcription, stress responses, and malignant transformation. They are found only in eukaryotes. In this study, I cloned and characterized 9 genes encoding nine members of zebrafish 14-3-3 family, designated theta (θ), zeta (ζ), beta1 (β1), beta2 (β2), beta3 (β3), eta (η), gamma (γ), epsilon1 (e1), epsilon2 (e2). Amino acid sequence alignment and phylogenetic tree analysis indicated that all zebrafish 14-3-3 members can be divided into three group. The ζ, θ, β1, β2 and β3 belong to Group I, while η and γ Group II. In addition, e1 and e2 are the third. Using whole-mount in situ hybridization to analyze the mRNA expression profile of zebrafish 14-3-3 family from 6 hpf to 120 hpf, the signal was found primarily in the brain while moderate in retina, gill and gut. Expression of GFP-fusion protein of each isoform of zebrafish 14-3-3 family by a neuron-specific HuC promoter in zebrafish embryos indicated that only 14-3-3-beta3 isoform could promote neurite outgrowth. A double mutant of 14-3-3-beta3 (R56A and R60A) abolished this effect. Coinjection of pHuC-14-3-3-beta3-HuC-GFP with protein phosphatase 2A (PP2A), Cdc42 dominant negative form (Cdc42-T17N), Rac1 dominant negative form (Rac1-T17N) and MAPKK inhibitor PD98059, respectively, reduced the neurite outgrowth induced by 14-3-3-beta3.
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