A novel plant nuclear gene encoding chloroplast ribosomal protein S9 has a transit peptide related to that of rice chloroplast ribosomal protein L12

Abstract

We have cloned a novel nuclear gene for a ribosomal protein of rice and Arabidopsis that is like the bacterial ribosomal protein S9. To determine the subcellular localization of the gene product, we fused the N-terminal region and green fluorescent protein and expressed it transiently in rice seedlings. Localized fluorescence was detectable only in chloroplasts, indicating that this nuclear gene encodes chloroplast ribosomal protein S9. The N-terminal region of rice ribosomal protein S9 was found to have a high sequence similarity to the transit peptide region of the rice chloroplast ribosomal protein L12, suggesting that these transit peptides have a common lineage.z 1999 Federation of European Biochemical Societies.

Mendel's green cotyledon gene encodes a positive regulator of the chlorophyll-degrading pathway

“…10). The signals for both constructs were colocalized with that of the red fluorescent protein (RFP) fused to the transit peptide from the plastid-localizing rice S9 ribosomal protein (18), suggesting that both PsSGR JI4 and PsSGR JI2775 are localized in plastids, and the two amino acid substitutions in the PsSGR JI2775 transit peptide are not responsible for the staygreen phenotype of JI2775. The two-amino acid insertion is located in a relatively conserved region of the SGR proteins, and there is no length difference in this region even between dicotyledous and monocotyledous SGR proteins (SI Fig.…”

“…A plastidlocalized RFP construct with the transit peptide from rice ribosomal protein S9 is the RFP derivative of OsPRS9TP-GFP (18). PsSGR JI2775 -GFP and PsSGR JI4 -GFP were constructed by insertion of the full-length PsSGR CDS fragment into the XbaI-BamHI site of pJ4-GFP (29).…”

Frequent fusion and fission of plant mitochondria with unequal nucleoid distribution

“…The coding region of Kaede was subcloned from the plasmid pKaede (Amalgaam, Tokyo) into the general plant expression vector with the CaMV35S promoter and the NOS terminator. The N-terminal 36 aa of the Arabidopsis mitochondrial ATPase ␦ subunit (21) and the N-terminal 98 aa of rice chloroplast ribosomal protein S9 (22) were subcloned into the Kaede plasmid as presequence and transit peptides, respectively. The PTS-1 peroxisome localization signal [Cterminal SKL sequence (23)] was generated from the Kaede expression vector by using a QuikChange XL Site-Directed Mutagenesis Kit (Stratagene).…”

The gene for alternative oxidase-2(AOX2) from Arabidopsis thaliana consists of five exons unlike other AOX genes and is transcribed at an early stage during germination.

“…Tobacco BY-2 cells were cultured for 36 hours after transformation in the dark. The transformed cells were then observed at × 40 magnification with a confocal laser scanning microscopy as described by Arimura et al (1999). To visualize localization of mitochondria in tobacco BY-2 cells, the cells were treated with 500 nM MitoTracker Orange CM-H 2 TMRos (Molecular Probes, Eugene, OR, USA), a mitochondrial-specific dye, for 30 min.…”

“…To examine this possibility, we constructed an AOX2pre-synthetic green fluorescent protein (GFP) fusion protein. GFP has been shown to be useful as a vital marker for the analysis of trafficking and subcellular localization of proteins (Chiu et al, 1996;Köhler et al, 1997;Arimura et al, 1999). The plasmid, termed AOX2pre-GFP, was constructed by fusing codons (coding for amino acid residues 1-72) of the predicted AOX2 precursor protein to the coding sequence of GFP (Fig.…”