The Arabidopsis thaliana K+ channel KAT1 has been suggested to have a key role in mediating the aperture of stomata pores on the surface of plant leaves. Although the activity of KAT1 is thought to be regulated by phosphorylation, the endogenous pathway and the primary target site for this modification remained unknown. In the present study, we have demonstrated that the C-terminal region of KAT1 acts as a phosphorylation target for the Arabidopsis calcium-independent ABA (abscisic acid)-activated protein kinase SnRK2.6 (Snf1-related protein kinase 2.6). This was confirmed by LC-MS/MS (liquid chromatography tandem MS) analysis, which showed that Thr306 and Thr308 of KAT1 were modified by phosphorylation. The role of these specific residues was examined by single point mutations and measurement of KAT1 channel activities in Xenopus oocyte and yeast systems. Modification of Thr308 had minimal effect on KAT1 activity. On the other hand, modification of Thr306 reduced the K+ transport uptake activity of KAT1 in both systems, indicating that Thr306 is responsible for the functional regulation of KAT1. These results suggest that negative regulation of KAT1 activity, required for stomatal closure, probably occurs by phosphorylation of KAT1 Thr306 by the stress-activated endogenous SnRK2.6 protein kinase.
;We characterized three Arabidopsis genes, AtpOMT1, AtpDCT1 and AtpDCT2, localized on chromosome 5 and homologous to spinach chloroplastic 2-oxoglutarate/malate transporter (OMT) gene. The yeast-expressed recombinant AtpOMT1 protein transported malate and 2-oxoglutarate but not glutamate. By contrast, the recombinant AtpDCT1 protein transported 2-oxoglutarate and glutamate at similar affinities in exchange for malate. These findings suggested that AtpOMT1 is OMT and AtpDCT1 is a general dicarboxylate transporter (DCT). The recombinant proteins could also transport oxaloacetate at the same binding sites for dicarboxylates. In particular, the AtpOMT1 had a K m value for oxaloacetate one order of magnitude lower than those for malate and 2-oxoglutarate. Although the transcripts for the three genes were accumulated in all tissues examined, the expression of the genes in leaf tissues was light inducible. The expression of the three genes was also induced by nitrate supplement but the induction was most prominent and transient in AtpOMT1 similar to nitrate reductase gene. These findings lead to a proposition that AtpOMT1 functions as an oxaloacetate transporter in the malate-oxaloacetate shuttle across chloroplast membranes. We identified T-DNA insertional mutants of AtpOMT1 and AtpDCT1. Although the AtpOMT1 mutants could grow normally in normal air, the AtpDCT1 mutants were non-viable under the same conditions. The AtpDCT1 mutants were able to grow under the high CO 2 condition to suppress photorespiration. These findings suggested that at least AtpDCT1 is a necessary component for photorespiratory nitrogen recycling.
We report the first study of sperm whale-fall ecosystems, based on mass sinking of whale carcasses at shelf depths in the northwest Pacific. We conducted three observations over a 2-year period on replicate sperm-whale carcasses implanted at depths of 219-254 m off the southern part of Japan from July 2003 to August 2005. The study was made possible by a mass stranding of sperm whales in January 2002, and the subsequent sinking of 12 carcasses in the waters off Cape Nomamisaki. Dense aggregations of unique chemosynthesis-based fauna had formed around the whale carcasses after 18 months (July 2003). The mytilid mussel Adipicola pacifica was the most abundant macrofaunal species and covered most of the exposed bone surfaces. The general composition of the fauna was similar to that of deep-water reducing habitats, but none of the species appearing in this study has been found at hydrothermal vents, cold seeps or deep-water whale falls. A new species of lancelet, which was the first record of the subphylum Cephalochordata from reducing environments, a new species of Osedax; a rarely encountered benthic ctenophore, and a rare gastropod species were discovered at this sperm whale-fall site. Benthic communities were similar across all the carcasses studied, although the body sizes of the whales were very different. The succession of epifaunal communities was relatively rapid and the sulphophilic stage was considerably shorter than that of other known whale falls.
Fatty replacement is more severe in the anterior aspect of the head of the pancreas. The posterior aspect of the head of the pancreas and the area around the CBD tended to be spared.
SUMMARYTransport of dicarboxylates across the chloroplast envelope plays an important role in transferring carbon skeletons to the nitrogen assimilation pathway and exporting reducing equivalent to the cytosol to prevent photo-inhibition (the malate valve). It was previously shown that the Arabidopsis plastidic 2-oxoglutarate/ malate transporter (AtpOMT1) and the general dicarboxylate transporter (AtpDCT1) play crucial roles at the interface between carbon and nitrogen metabolism. However, based on the in vitro transport properties of the recombinant transporters, it was hypothesized that AtpOMT1 might play a dual role, also functioning as an oxaloacetate/malate transporter, which is a crucial but currently unidentified component of the chloroplast malate valve. Here, we test this hypothesis using Arabidopsis T-DNA insertional mutants of AtpOMT1. Transport studies revealed a dramatically reduced rate of oxaloacetate uptake into chloroplasts isolated from the knockout plant. CO 2 -dependent O 2 evolution assays showed that cytosolic oxaloacetate is efficiently transported into chloroplasts mainly by AtpOMT1, and supported the absence of additional oxaloacetate transporters. These findings strongly indicate that the high-affinity oxaloacetate transporter in Arabidopsis chloroplasts is AtpOMT1. Further, the knockout plants showed enhanced photo-inhibition under high light due to greater accumulation of reducing equivalents in the stroma, indicating malfunction of the malate valve in the knockout plants. The knockout mutant showed a phenotype consistent with reductions in 2-oxoglutarate transport, glutamine synthetase/glutamate synthase activity, subsequent amino acid biosynthesis and photorespiration. Our results demonstrate that AtpOMT1 acts bi-functionally as an oxaloacetate/malate transporter in the malate valve and as a 2-oxoglutarate/malate transporter mediating carbon/nitrogen metabolism.
Recent culture-independent molecular analyses have shown the diversity and ecological importance of microbial eukaryotes (protists) in various marine environments. In the present study we directly extracted DNA from anoxic sediment near active fumaroles on a submarine caldera floor at a depth of 200 m and constructed genetic libraries of PCR-amplified eukaryotic small-subunit (SSU) rDNA. By sequencing cloned SSU rDNA of the libraries and their phylogenetic analyses, it was shown that most sequences have affiliations with known major lineages of eukaryotes (Cercozoa, Alveolata, stramenopiles and Opisthokonta). In particular, some sequences were closely related to those of representatives of eukaryotic parasites, such as Phagomyxa and Cryothecomonas of Cercozoa, Pirsonia of stramenopiles and Ichthyosporea of Opisthokonta, although it is not clear whether the organisms occur in free-living or parasitic forms. In addition, other sequences did not seem to be related to any described eukaryotic lineages suggesting the existence of novel eukaryotes at a high-taxonomic level in the sediment. The community composition of microbial eukaryotes in the sediment we surveyed was different overall from those of other anoxic marine environments previously investigated.
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