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Y Dokiya

Publications and source records attributed to Y Dokiya.

5 recordsLinked to original sources

Dual targeting of phage-type RNA polymerase to both mitochondria and plastids is due to alternative translation initiation in single transcripts.

We isolated and sequenced a nuclear gene and cDNA encoding a bacteriophage T7-type RNA polymerase, NsRpoT-B, from Nicotiana sylvestris. The gene, NsRpoT-B, consists of 19 exons and 18 introns and encodes a polypeptide of 1020 amino acid residues. The predicted NsRpoT-B protein shows 71% amino acid identity with NsRpoT-A which is a mitochondrial protein. Quantitative RT-PCR revealed that steady-state NsRpoT-B mRNA accumulation is highest in the mature leaves and lowest in the cotyledons. Transient expression assays in protoplasts from N. sylvestris leaves demonstrated that the putative N-terminal transit peptide of NsRpoT-B encodes dual targeting signals directing the protein into mitochondria and plastids. This strongly suggests that NsRpoT-B functions as an RNA polymerase transcribing genes from two different plant organelle genomes. NsRpoT-B transcripts have two potential translation initiation codons. An in vitro translation assay indicated that a chimeric mRNA encoding the N-terminal NsRpoT-B fused to an sGFP produced two polypeptides translated from the first and second initiation codons. This implies that the dual targeting of NsRpoT-B protein is regulated, in part, at the level of translation. We have designated this protein NsRpoTpm.

Alternative Splicing↗

Genomic organization and organ-specific expression of a nuclear gene encoding phage-type RNA polymerase in Nicotiana sylvestris.

We have isolated and sequenced a nuclear gene and cDNA encoding bacteriophage T7-type single subunit RNA polymerase, NsRpoT-A, from Nicotiana sylvestris. NsRpoT-A consists of 19 exons and 18 introns; the first intron is 17 kb, the longest yet identified in a plant gene. Genomic Southern analysis indicated that N. sylvestris contains a small family of NsRpoT genes. Quantitative RT-PCR revealed that steady-state mRNA levels are highest in the leaves and lowest in the cotyledons. Phylogenetic analysis of NsRpoT-A and the RpoT proteins of other plant species suggested that NsRpoT-A is a mitochondrial protein. The TargetP program predicted localization of the NsRpoT-A gene product to the mitochondria. Using a transient expression assay and protoplasts from N. sylvestris mesophyll cells, we clearly demonstrated that the N-terminal sequence of NsRpoT-A targets the protein to the mitochondria. We therefore named this protein NsRpoTm.

Amino Acid Sequence↗

Acid fog damage of beech at Hinokiboramaru-Tanzawa, Japan.

Decline of Japanese beech forest has been obvious at the Tanzawa Mountains near Tokyo. Observations on fog, precipitation and stem flow were performed during June-October, 1994 on both, NE (with healthy beech) and S-SE (with unhealthy beech) slopes, collecting samples biweekly. Chemical species were determined by ion-chromatography and pH meter. The results show that the pH of the precipitation was between 4.4-5.2, with no clear differences in the concentrations of chemical species between both sites and seasons. The pH of the fog water was high from May to the middle of August and lower from mid-August to October at both sites. The concentrations of most chemical species were higher in fog samples than precipitation samples. The concentrations of potassium and magnesium ions in the stem flow of an unhealthy tree on the S-SE slope were higher than those on the NE slope. In June-October, 1995, ten pairs of event fog water samples were obtained with indicating that the pH was below 4.0 for five pairs of event fog water samples. Lower pH value was obtained on the S-SE slope than on the NE slope in every event. Concentrations of most chemical species were always higher in the fog samples from the S-SE slope than those from the NE slope. The equivalent ratio of NO3-/nssSO4(2-) was higher in the fogs of higher acidity. From the result of atmospheric current analysis using a model, it was suggested that the polluted air over the Tokyo Metropolitan Area affected the fog acidity directly and also indirectly, according to the pressure patterns.

Acid Rain↗