Structure and differential expression of two distinct genes encoding the chloroplast elongation factor Tu in tobacco

Abstract

We have isolated two nuclear genes, tufA and tufB, encoding chloroplast EF-Tu from a tobacco (Nicotiana sylvestris) genomic library. The tufA gene encodes a polypeptide of 478 amino-acid residues, consisting of a putative transit peptide of 70 residues and a mature EF-TuA of 408 residues. The tufB gene codes for a precursor proteins of 485 residues, containing a transit peptide of 77 residues and a mature EF-TuB of 408 residues. No introns were found in either gene. The sequence similarity within the coding regions of the two genes is 84.3% for nucleotides and 89.7% for amino acids. Multiple 5' ends of transcripts were observed for both tuf genes. Northern analysis revealed that the EF-Tu mRNA accumulated at least 30-fold more in leaf than in root tissue. Ribonuclease protection assays using gene-specific probes showed that the level of tufB mRNA is three-fold higher than that of tufA mRNA in leaves but in roots the tufB mRNA levels is less than half that of tufA mRNA. The relative amount of tufB mRNA is 30-fold higher in leaves than in roots whereas tufA messages are only five-fold higher in leaves. These data suggest that expression of both tuf genes is differentially regulated according to tissue and plastid type.