cDNA structure, expression and nucleic acid-binding properties of three RNA-binding proteins in tobacco: occurence of tissue-specific alternative splicing

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Abstract
Three cDNAs encoding RNA-binding proteins were isolated from a tobacco (Nlcotiana sylvestris) cDNA library. The predicted proteins (RGP-1) are homologous to each other and consist of a consensus-sequence type RNA-binding domain of 80 amlno acids in the N–terminal half and a glycine-rlch domain of 61-78 amino acids in the C-termlnal half. Nucleic acid-binding assay using the In vitro synthesized RGP-1 protein confirmed that it is an RNA-binding protein. Based on its strong affinity for poly(G) and poly(U), the RGP-1 proteins are suggested to bind specifically to G and/or U rich sequences. All three genes are expressed in leaves, roots, flowers and cultured cells, however, the substantial amount of pre-mRNAs are accumulated especially in roots. Sequence analysis and ribonuclease protection assay Indicated that significant amounts of alternatively spliced mRNAs, which are produced by differential selection of 5' splice sites, are also present in various tissues. Tissue-specific alternative splicing was found in two of the three genes. The alternatively spliced mRNAs are also detected in polysomal fractions and are suggested to produce truncated polypeptides. A possible role of this alternative splicing is discussed.