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Identification of a Functional Splice Variant of 14-3-3E1 in Rainbow Trout

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Abstract

The 14-3-3 protein family is a family of regulatory proteins involved in diverse cellular processes. The presence of 14-3-3 isoforms and the diversity of cellular processes regulated by 14-3-3 isoforms suggest functional specificity of the isoforms. In this study, we report the identification and characterization of a new isoform of the rainbow trout 14-3-3E1 gene generated by alternative splicing. The new isoform contains an insertion of 48 nucleotides (from intron 5) in the coding region of 14-3-3E1 which results in the introduction of a premature stop codon between exon 5 and exon 6. Thus, the alternatively spliced form of 14-3-3E1 (14-3-3E1∆C17) lacks 17 amino acid residues at the C terminus encoded by the last exon (exon 6). Reverse-transcription polymerase chain reaction analysis revealed that the wild-type 14-3-3E1 (14-3-3E1wt) is ubiquitously expressed, while 14-3-3E1∆C17 shows tissue-specific as well as stage-specific expression during ovarian development and early embryogenesis. Analysis by yeast two-hybrid system demonstrated that 14-3-3E1∆17 interacts with a number of proteins including ATP synthase, ankyrin repeat domain 13b, cytochrome c subunit VIa, cytochrome c subunit VIb, 60S ribosomal protein L34, solute carrier family 17 member 6 (SLC17A6), troponin I, and an unknown protein. Although all of these proteins except for SLC17A6 also interact with 14-3-3E1wt, 14-3-3E1∆17 appears to have higher binding affinity with these proteins than 14-3-3E1wt. These findings suggest that alternative splicing affects the function and tissue-specific expression of 14-3-3E1.

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Acknowledgements

This investigation was supported by Hatch Funds (project no. 427) of the West Virginia Agricultural Experiment Station and USDA/ARS Cooperative Agreement No. 58-1930-5-537. It is published with the approval of the Director of the West Virginia Agriculture and Forestry Experiment Station as scientific paper no. 3039.

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Authors and Affiliations

  1. Division of Animal and Nutritional Sciences, West Virginia University, Morgantown, WV, 26506, USA

    Warapond Wanna & Jianbo Yao

  2. Center of Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, 90112, Thailand

    Warapond Wanna

  3. National Center for Cool and Cold Water Aquaculture, Kearneysville, WV, 25430, USA

    Caird E. Rexroad III

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  1. Warapond Wanna
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  2. Caird E. Rexroad III
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Correspondence to Jianbo Yao.

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Wanna, W., Rexroad, C.E. & Yao, J. Identification of a Functional Splice Variant of 14-3-3E1 in Rainbow Trout. Mar Biotechnol 12, 70–80 (2010). https://doi.org/10.1007/s10126-009-9201-6

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