A single nuclear transcript encoding mitochondrial RPS14 and SDHB of rice is processed by alternative splicing: common use of the same mitochondrial targeting signal for different proteins.
The rice mitochondrial genome has a sequence homologous to the gene for ribosomal protein S14 (rps14), but the coding sequence is interrupted by internal stop codons. A functional rps14 gene was isolated from the rice nuclear genome, suggesting a gene-transfer event from the mitochondrion to the nucleus. The nuclear rps14 gene encodes a long N-terminal extension showing significant similarity to a part of mitochondrial succinate dehydrogenase subunit B (SDHB) protein from human and a malarial parasite (Plasmodium falciparum). Isolation of a functional rice sdhB cDNA and subsequent sequence comparison to the nuclear rps14 indicate that the 5' portions of the two cDNAs are identical. The sdhB genomic sequence shows that the SDHB-coding region is divided into two exons. Surprisingly, the RPS14-coding region is located between the two exons. DNA gel blot analysis indicates that both sdhB and rps14 are present at a single locus in the rice nucleus. These findings strongly suggest that the two gene transcripts result from a single mRNA precursor by alternative splicing. Protein blot analysis shows that the size of the mature RPS14 is 16.5 kDa, suggesting removal of the N-terminal 22.6-kDa peptide region. Considering that the rice mitochondrial genome lacks the sdhB gene but contains the rps14-related sequence, transfer of the sdhB gene seems to have occurred before the transfer of the rps14 gene. The migration of the mitochondrial rps14 sequence into the already existing sdhB gene could bestow the capacity for nuclear expression and mitochondrial targeting.
Kubo
N
N
Faculty of Horticulture, Chiba University, Matsudo 648, Matsudo, Chiba 271-0092, Japan.
Harada
K
K
Hirai
A
A
Kadowaki
K
K
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics*; D018076:DNA, Complementary; D004272:DNA, Mitochondrial / Q000235:genetics*; D005091:Exons; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D018745:Genome, Plant; D006801:Humans; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics; D021122:Protein Subunits; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013385:Succinate Dehydrogenase / Q000235:genetics
D018076:DNA, Complementary; D004272:DNA, Mitochondrial; D007506:Iron-Sulfur Proteins; D021122:Protein Subunits; C108095:SDHB protein, human; D013385:Succinate Dehydrogenase