Structural Studies of Inositol 1,4,5-Trisphosphate Receptor: COUPLING LIGAND BINDING TO CHANNEL GATING*

The three isoforms of the inositol 1,4,5-trisphosphate receptor (IP₃R) exhibit distinct IP₃ sensitivities and cooperativities in calcium (Ca²⁺) channel function. The determinants underlying this isoform-specific channel gating mechanism have been localized to the N-terminal suppressor region of IP₃R. We determined the 1.9 Å crystal structure of the suppressor domain from type 3 IP₃R (IP₃R3_SUP, amino acids 1–224) and revealed structural features contributing to isoform-specific functionality of IP₃R by comparing it with our previously determined structure of the type 1 suppressor domain (IP₃R1_SUP). The molecular surface known to associate with the ligand binding domain (amino acids 224–604) showed marked differences between IP₃R3_SUP and IP₃R1_SUP. Our NMR and biochemical studies showed that three spatially clustered residues (Glu-20, Tyr-167, and Ser-217 in IP₃R1 and Glu-19, Trp-168, and Ser-218 in IP₃R3) within the N-terminal suppressor domains of IP₃R1_SUP and IP₃R3_SUP interact directly with their respective C-terminal fragments. Together with the accompanying paper (Yamazaki, H., Chan, J., Ikura, M., Michikawa, T., and Mikoshiba, K. (2010) J. Biol. Chem. 285, 36081–36091), we demonstrate that the single aromatic residue in this region (Tyr-167 in IP₃R1 and Trp-168 in IP₃R3) plays a critical role in the coupling between ligand binding and channel gating.     

Effect of nutritional condition on photosymbiotic consortium of cultured <Emphasis Type="Italic">Globigerinoides sacculifer</Emphasis> (Rhizaria,Foraminifera)

Several foraminifers found in warm and low-nutrient ocean surface water have photosynthetic algae as endosymbionts (photosymbiosis). To understand the trophic interactions, we studied Globigerinoides sacculifer, a spinose planktic foraminifer that has a dinoflagellate endosymbiont. We controlled two nutritional factors, feeding and inorganic nutrients in the seawater. The growth of the host and the symbionts and the photophysiological parameters were monitored under four experimental conditions. The results demonstrated that the holobionts primarily relied on phagotrophy for growth. The foraminifers grew considerably, and the chlorophyll a content per foraminifer, which is an indicator of the symbiont population, increased in the fed groups, but not in the unfed groups. The nutrient-rich seawater used for some of the cultures made no difference in either the growth or photophysiology of the holobionts. These observations indicated that the symbionts mainly utilized metabolites from the hosts for photosynthesis rather than inorganic nutrients in the seawater. Additionally, we observed that the symbionts in the starved hosts maintained their photosynthetic capability for at least 12 days, and that the hosts maintained at least some symbionts until gametogenesis was achieved. This suggests that the hosts have to retain the symbionts as an energy source for reproduction. The symbionts may also play an indispensable role in the metabolic activities of the hosts including waste transport or essential compound synthesis. Overall, our results revealed a novel mode of photosymbiosis in planktic foraminifers which contrasts with that found in benthic photosymbiotic foraminifers and corals.     

A lemon myrtle extract inhibits glucosyltransferases activity of Streptococcus mutans

Streptococcus mutans is a bacterium found in human oral biofilms (dental plaques) that is associated with the development of dental caries. Glucosyltransferases (GTFs) are key enzymes involved in dental plaque formation, and compounds that inhibit their activities may prevent dental caries. We developed a screening system for GTF-inhibitory activities, and used it to profile 44 types of herbal tea extracts. Lemon myrtle (Backhousia citriodora) extract exhibited the highest GTF-inhibitory activity, with an IC₅₀ for GTF in solution of 0.14 mg mL^?1. Furthermore, lemon myrtle extracts had the third-highest polyphenol content of all tested extracts, and strongly inhibited S. mutans biofilm. Interestingly, lemon myrtle extracts did not inhibit cell growth.     

Drought stress during soybean seed filling affects storage compounds through regulation of lipid and protein metabolism

Soybean seeds have high lipid and protein contents. Adverse environmental conditions restrict seed yield and quality. We examined the changes in storage compounds caused by drought stress from R5 stage (beginning seed growth stage). Under drought stress, contents of lipid in seed were remarkably low compared to control at 24 and 29 days after treatment. Protein contents in seed were immediately decreased after water deficit treatment. On the other hand, soluble sugar contents in seed were increased by drought stress. Drought stress decreased the expression of genes involved in lipid biosynthesis (PK, BCCP2, and KAS1) and increased the genes expression involved in lipid degradation (ACX2, MS, and PEPCK). These results suggest that the increasing of sugar content in seed under drought stress was complemented by degradation of lipids. The expressions of genes encoding storage protein (Gy4 and β-conglycinin) were also decreased by drought stress. This study showed how drought stress during seed filling affects seed quality, especially lipid and protein contents, that may facilitate further research on seed storage compounds metabolism under environmental stresses.     

Role of transpiration from fruits in phloem transport and fruit growth in tomato fruits

Sink activity of fruits had been suggested to vary depending on transpiration of fruits. In this study, the effect of transpiration on dry matter accumulation was evaluated in tomato ( Lycopersicon esculentum Mill.). Fruits of cv. Saturn at 14 days after anthesis were enclosed in chambers and aerated with dried (<15% RH) or moistened (>90% RH) air. These treatments did not cause any significant differences in fruit fresh weight, dry weight, percentage of dry matter, and concentration of soluble sugars within 5 days of the treatment, or the import of ¹⁴C within 18 h after the application of ¹⁴CO₂ to the source leaves. However, displacement transducer measurement of each fruit showed a 40% reduction in growth rate in response to exchange of moistened air with dried air. When fruits of cv. Momotaro were exposed to transpiration treatments from the beginning of visible fruit enlargement until the ripening stage, the fruits exhibited 20% reduction in growth and lower accumulation of dry matter at harvest following treatment with dried air. These results suggested that higher transpiration reduced both water accumulation and dry matter accumulation. In contrast, when fruit growth was mechanically restricted by enclosing the fruits in a chamber packed with glass beads, and dried or moistened air was passed through the spaces between the glass beads, fruits exhibited higher dry matter accumulation under dried air treatment conditions. The results show that only under artificial conditions would transpiration of fruits potentially drive carbohydrate transport; it does not serve as a limiting step of carbohydrate transport to tomato fruits under normal circumstances.     

Land abandonment and changes in snow cover period accelerate range expansions of sika deer

Ongoing climate change and land‐use change have the potential to substantially alter the distribution of large herbivores. This may result in drastic changes in ecosystems by changing plant–herbivore interactions. Here, we developed a model explaining sika deer persistence and colonization between 25 years in terms of neighborhood occupancy and habitat suitability. We used climatic, land‐use, and topographic variables to calculate the habitat suitability and evaluated the contributions of the variables to past range changes of sika deer. We used this model to predict the changes in the range of sika deer over the next 100 years under four scenario groups with the combination of land‐use change and climate change. Our results showed that both climate change and land‐use change had affected the range of sika deer in the past 25 years. Habitat suitability increased in northern or mountainous regions, which account for 71.6% of Japan, in line with a decrease in the snow cover period. Habitat suitability decreased in suburban areas, which account for 28.4% of Japan, corresponding to land‐use changes related to urbanization. In the next 100 years, the decrease in snow cover period and the increase in land abandonment were predicted to accelerate the range expansion of sika deer. Comparison of these two driving factors revealed that climate change will contribute more to range expansion, particularly from the 2070s onward. In scenarios that assumed the influence of both climate change and land‐use change, the total sika deer range increased by between +4.6% and +11.9% from the baseline scenario. Climate change and land‐use change will require additional efforts for future management of sika deer, particularly in the long term.     

Relationship between maternal environment and DNA methylation patterns of estrogen receptor alpha in wild Eastern Bluebird (Sialia sialis) nestlings: a pilot study

There is mounting evidence that, across taxa, females breeding in competitive environments tend to allocate more testosterone to their offspring prenatally and these offspring typically have more aggressive and faster‐growing phenotypes. To date, no study has determined the mechanisms mediating this maternal effect's influence on offspring phenotype. However, levels of estrogen receptor alpha (ERα) gene expression are linked to differences in early growth and aggression; thus, maternal hormones may alter gene regulation, perhaps via DNA methylation, of ERα in offspring during prenatal development. We performed a pilot study to examine natural variation in testosterone allocation to offspring through egg yolks in wild Eastern Bluebirds (Sialia sialis) in varying breeding densities and percent DNA methylation of CG dinucleotides in the ERα promoter in offspring brain regions associated with growth and behavior. We hypothesized that breeding density would be positively correlated with yolk testosterone, and prenatal exposure to maternal‐derived yolk testosterone would be associated with greater offspring growth and decreased ERα promoter methylation. Yolk testosterone concentration was positively correlated with breeding density, nestling growth rate, and percent DNA methylation of one out of five investigated CpG sites (site 3) in the diencephalon ERα promoter, but none in the telencephalon (n = 10). Percent DNA methylation of diencephalon CpG site 3 was positively correlated with growth rate. These data suggest a possible role for epigenetics in mediating the effects of the maternal environment on offspring phenotype. Experimentally examining this mechanism with a larger sample size in future studies may help elucidate a prominent way in which animals respond to their environment. Further, by determining the mechanisms that mediate maternal effects, we can begin to understand the potential for the heritability of these mechanisms and the impact that maternal effects are capable of producing at an evolutionary scale.     

Isolation and Properties of Suppressor-Sensitive Mutants of Bacillus subtilis Bacteriophage SPO2

Suppressor-sensitive mutants of Bacillus subtilis phage SPO2 were isolated and classified into 18 cistrons. A linear genetic map was constructed. The genes controlling early functions of phage development were clustered near the CI gene which may produce phage repressor.