Cloning and expression analyses of cDNA encoding an ADP-ribosylation factor from wheat: tissue-specific expression of...

We isolated and characterized a cDNA clone encoding a small GTP-binding protein, which has a high similarity to mammalian ADP-ribosylation factor (ARF), from a cDNA library prepared from immature spikes of wheat (Triticum aestivum L.). The cDNA contained an open reading frame that encodes a polypeptide of 181 amino acids with a calculated molecular mass of 20.7 kDa. The deduced amino acid sequence showed a quite high homology to known ARFs from other organisms. In particular, the wheat ARF was completely identical to the rice ARF1. Genomic Southern hybridization suggested that wheat ARF is encoded by at least two or three copies of ARF genes. Northern analyses showed that the accumulation of the ARF transcripts was nearly constant throughout various environmental stresses in both shoots and roots. However the RNA transcript was preferentially expressed in roots rather than in shoots. A similar expression pattern was also observed at the protein level by Western analysis. The relative abundance of the ARF proteins in root and flower tissues may indicate a high level of vesicular transporting activity in the roots and flowers of wheat plants.