Molecular genetic and chemotaxonomic characterization of the terrestrial cyanobacterium Nostoc commune and its neighboring species.

The phylogeny of the terrestrial cyanobacterium Nostoc commune and its neighboring Nostoc species was studied using molecular genetic and chemotaxonomic approaches. At least eight genotypes of N. commune were characterized by the differences among 16S rRNA gene sequences and the petH gene encoding ferredoxin-NADP⁺ oxidoreductase and by random amplified polymorphic DNA analysis. The genotypes of N. commune were distributed in Japan without regional specificity. The nrtP gene encoding NrtP-type nitrate/nitrite permease was widely distributed in the genus Nostoc, suggesting that the occurrence of the nrtP gene can be one of the characteristic features that separate cyanobacteria into two groups. The wspA gene encoding a 36-kDa water stress protein was only found in N. commune and Nostoc verrucosum, suggesting that these Nostoc species that form massive colonies with extracellular polysaccharides can be exclusively characterized by the occurrence of the wspA gene. Fifteen species of Nostoc and Anabaena were investigated by comparing their carotenoid composition. Three groups with distinct patterns of carotenoids were related to the phylogenic tree constructed on the basis of 16S rRNA sequences. Nostoc commune and Nostoc punctiforme were clustered in one monophyletic group and characterized by the occurrence of nostoxanthin, canthaxanthin, and myxol glycosides.

Group 3 sigma factors in the marine cyanobacterium Synechococcus sp. strain PCC 7002 are required for growth at low temperature.

Three genes, sigF, sigG and sigH, encoding group 3 sigma factors have been cloned and characterized in the marine cyanobacterium Synechococcus sp. strain PCC 7002. The sigF gene product was similar to sigma factors involved in general stress response and sporulation in other organisms, and the sigG and sigH gene products were similar to extracytoplasmic function (ECF) sigma factors. The sigG and sigH genes were associated with the putative regulatory genes and the sizes of transcripts for sigG and sigH genes were large enough to be cotranscribed with the associated downstream genes. The sigG downstream gene was designated sapG (sigG-associated protein), and yeast two-hybrid analysis demonstrated that SigG and SapG interact when produced in yeast cells. Null mutants of these three group 3 sigma factor genes were created by interposon mutagenesis. The growth of the sigF mutant strain was much slower than the wild-type strain at 15 degrees C, although the growth rates at 22 degrees C and 38 degrees C were identical to those of the wild-type strain. The sigG mutant could not grow continuously at 22 degrees C, and no growth occurred at 15 degrees C. Since SigG and SapG interact in yeast cells and the sigG and sapG mutants showed a similar growth phenotype, SapG is likely to be a regulatory protein for SigG involved in the same pathway in transcriptional regulation in this cyanobacterium.

Selective scavenging property of the indole moiety for the nitrating species of peroxynitrite.

The inhibitory effect on tyrosine nitration and oxidation of peroxynitrite was evaluated for more than 40 reagents including natural and synthetic compounds, and the inhibiting efficiency of each compound for nitration was compared with that for oxidation, to characterize its property as a peroxynitrite scavenger. In the presence of various concentrations of testing compounds, the nitrating and oxidizing activities were measured by monitoring the formation of 3-nitrotyrosine and dityrosine with an HPLC-UV-fluorescence detector. The IC(50) values for nitration and oxidation were determined, and the ratio of these two IC(50) values was calculated for each compound. Although the IC(50) values varied from compound to compound, it was revealed that the ratio of two IC(50) values (IC(50) for oxidation/IC(50) for nitration) was 1 in almost all the compounds tested, except five indole derivatives (L-tryptophan, melatonin, 5-methoxytryptamine, tryptamine, and tetrahydro-beta-carboline) and one synthetic selenium-containing compound ((2R,3R,4S)-2-amino-3,4-dihydroxy-5-phenylselenopentan-1-ol, ADPP). The indole derivatives showed a specific inhibitory effect on tyrosine nitration without affecting the oxidation. ADPP was confirmed to have a preferable inhibitory activity for tyrosine oxidation. It was suggested that compounds showing an IC(50) value ratio of 1 scavenged the common species for nitration and oxidation, while the indole derivatives and ADPP preferably scavenged the nitrating and oxidizing species, respectively. From a stopped flow study, it was also revealed that the nitrotyrosine formation was relatively slow, unlike an OH radical reaction. These results imply that the peroxynirite reaction at least partly proceeds through specific species for nitration.

Radioprotective effect of chitosan in sub-lethally X-ray irradiated mice.

The radioprotective effect of chitosan was studied in mice following whole-body X-ray irradiation. C3H/He mice were exposed to 7 Gy, and their survival rates were examined. The survival rates of chitosan-diet mice were about 20% higher than those of mice on a standard diet, and the rates dropped sharply to a plateau at day 10 after X-ray irradiation. The chitosan-diet mice had an increased weight ratio of spleen to body within the experimental period. The leukocyte, thrombocyte, and erythrocyte counts as well as the hematocrit and hemoglobin levels were recovered significantly and more rapidly in the chitosan-diet mice than the standard-diet mice at day 14 after irradiation. The scavenging abilities of chitosan were evaluated by the ESR spin-trapping method. These observations suggested that chitosan led to hematopoetic activation and leukocytogenesis in mice after sub-lethal dose irradiation, and that the biological response might be caused by radical trapping or scavenging.