Evidence for transit peptide acquisition through duplication and subsequent frameshift mutation of a preexisting protein gene in rice.
Many proteins synthesized in the cytosol are delivered to their appropriate compartments in the cell by specific targeting signals. Here, we provide new insight into the generation of the chloroplast-targeting signal (called the transit peptide) in rice. First, we identified the mitochondrial ribosomal protein L13 (mt rpl13) gene on chromosome 5. Downstream of the gene, we identified a DNA fragment of 266 bp: a segment within a duplication of mt rpl13. The duplicated region was transcribed and found to encode an open reading frame (ORF) of 160 amino acids (aa) (orf160). The orf160 gene comprises C-terminal 60 aa derived from the mt rpl13 gene and N-terminal 100 aa derived from another duplicated fragment of a pentatricopeptide repeat (ppr)564 gene that encodes 564 aa with ppr motifs on chromosome 1. Examination of the localization of the ORF160 protein tagged with green fluorescent protein (GFP) showed that it is targeted to the chloroplasts. As such, ORF160 clearly contains a transit peptide. Interestingly, this was translated from the alternative reading frame of the duplicated fragment of ppr564. To confirm this, the reading frame of the ppr564 gene was shifted according to that of the orf160 gene, and the frameshifted ppr564 sequence was fused to the gene for GFP. The expressed GFP-fused protein was also located in the chloroplasts. These results provide clear evidence for the generation of the transit peptide through duplication and subsequent frameshifting of a reading frame of a preexisting protein gene. We also demonstrate the importance of sequence redundancy and frameshift mutation in this evolutionary process.
Ueda
Minoru
M
Genetic Diversity Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.
Fujimoto
Masaru
M
Arimura
Shin-ichi
S
Tsutsumi
Nobuhiro
N
Kadowaki
Koh-ichi
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics; D002736:Chloroplasts / Q000235:genetics / Q000502:physiology; D004272:DNA, Mitochondrial / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics; D019143:Evolution, Molecular; D016368:Frameshift Mutation; D020440:Gene Duplication; D008954:Models, Biological; D008969:Molecular Sequence Data; D050506:Mutant Chimeric Proteins / Q000235:genetics / Q000502:physiology; D016366:Open Reading Frames / Q000235:genetics; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics*; D012269:Ribosomal Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D004272:DNA, Mitochondrial; D029721:Drosophila Proteins; D050506:Mutant Chimeric Proteins; D010940:Plant Proteins; D021382:Protein Sorting Signals; D012269:Ribosomal Proteins; C484179:RpL13 protein, Drosophila