Regulation of biosynthesis of aspartate family amino acids in the photosynthetic bacterium Rhodopseudomonas palustris

The four amino acids of the aspartate family (l-lysine, l-methionine, l-threonine, and l-isoleucine) are produced in bacteria by a branched biosynthetic pathway. Regulation of synthesis of early common intermediates and of carbon flow through distal branches of the pathway requires operation of a number of subtle feedback controls, which are integrated so as to ensure balanced synthesis of the several end products. Earlier studies with nonsulfur purple photosynthetic bacteria were instrumental in revealing the existence of alternative regulatory schemes, and in this communication we report on the control pattern of a representative of this physiological group not previously investigated, Rhodopseudomonas palustris. The results obtained from study of the properties of four key regulatory enzymes of the aspartate family pathway (-aspartokinase, homoserine dehydrogenase, homoserine kinase, and threonine deaminase) and of the effects of exogenous amino acids (i. e., the end products) on growth of the bacterium indicate that the control schema in Rps. palustris differs substantially from the schemes described for other Rhodopseudomonas species, but resembles the regulatory pattern observed in Rhodospirillum rubrum.Abbreviations A absorbancy - AK -aspartokinase - ASA aspartate -semialdehyde - DTT dithiothreitol - HS l-homoserine - HSDH homoserine dehydrogenase - HSK homoserine kinase - I l-isoleucine - KU Klett-Summerson photometer units - L l-lysine - M l-isoleucine - KU Klett-Summerson photometer units - L l-lysine - M l-methionine - ME -mercaptoethanol - PABA p-aminobenzoic acid - T l-threonine - TD threonine deaminase - RCV synthetic growth medium (see text) - YP agar medium containing 0.3% yeast extract, 0.3% peptone, and 1.5% agar - Y2T synthetic growth medium (see text)     

Energy expenditure and locomotor activity in mice selected for food intake adjusted for body weight

Summary The aim of this study was to examine the differences in physical activity and their contribution to differences in energy utilization in mice, selected either high or low for food intake, adjusted for body weight, which show correlated responses in lean content and metabolic rate. Simultaneous measurements of fasting metabolic rate and activity were made in lines of mice selected at either: a young age, 4-to 6-week food intake corrected for 4-week body weight; or an older age, 8- to 10-week food intake corrected for mean weight at 8 and 10 weeks of age. Correlated response in metabolic rate was found to have been accompanied by changes in locomotor activity near the ages at selection in both sets of lines. Activity, however, accounted for only a small proportion of variation in fasting heat production, generally less than 5%, although a highly positive correlation (r=0.63) between the two traits was found. It was concluded that selection for food intake adjusted for body weight has led to correlated response in physical activity. In consequence, mice selected in the upward direction expend some of the excess energy intake rather than assimilating it as body mass and are, therefore, slightly leaner than their counterparts selected in the downward direction.     

Effect of anoxia on the kinetic properties of pyruvate kinase and phosphofructokinase,and on glycogen phosphorylase activity in marine worms and earth worms

Summary The kinetic properties of PK and PFK were studied in aerobic versus 12-hours anoxic marine worms Hedistae(=Nereis) diversicolor and Diopatra neapolitana and earth worms Allolobophora calliginosa and Eisenia foetida. The total glycogen phosphorylase (a+b) activity and the percentage of active a form were also measured in the marine and earth worms under the same conditions. Anoxia exposure did not result in any significant changes of kinetic parameters of PK and total activities of glycogen phosphorylase from marine worms, but it altered the kinetic characteristics of PFK from H. diversicolor. Chromatographical studies showed that PK from both aerobic and anoxic marine worms is eluted from DEAE-cellulose as a single peak at 50 mM KCl. In contrast to marine worms, however, anoxia caused a marked change in kinetic properties of PK from both earth worms, resulting in a reduction of enzyme affinity for its substrate PEP. In addition, the enzyme existed in both earth worms in two distinct variants eluted from DEAE-cellulose column as peak I and peak II at 50 mM and 150 mM KCl, respectively. The ratio of enzyme units (peak I/peak II) was reduced significantly after 12 h of anoxia, indicating that these two peaks are interconvertible. Anoxia also caused a reduction of total glycogen phosphorylase activity in E. foetida and lowered the percentage of active a form of the enzyme by approximately 50% in both earth worms. Kinetic properties of PFK from both earth worms were not significantly affected by anoxia. However, their low Ka values for F-2,6-P₂ imply that this effector may play an important role in PFK control in earth worms under anoxia.Abbreviations F6P fructose-6-phosphate - FBP fructose-1,6-bisphosphate - F-2,6-P fructose-2,6-bisphosphate - PEP phosphoenoylpruvate - PFK 6-phosphofructo-1-kinase (E.C.2.7.1.11) - PK pyruvate kinase (E.C.2.7.1.40) - Pi inorganic phosphate - PMSF phenyl methylsulfonyl fluoride     

Regional Reductions of Transketolase in Thiamine-Deficient Rat Brain

Abstract: Thiamine deficiency impairs oxidative metabolism and causes metabolic encephalopathy. An early reduction in transketolase (TK) activity may be an important pathogenic event. To assess the role of TK, we have delineated the regional/cellular distribution of TK protein and mRNA in adult rat brain in pyrithiamine-induced thiamine deficiency. TK activity declined in both vulnerable and spared regions. Immunoblots showed a parallel reduction of TK protein. With a few exceptions, immunocytochemistry indicated an overall decline of TK immunoreactivity and the decrease was not specific to vulnerable areas. In contrast to the pronounced, general decline of TK protein, in situ hybridization revealed a regional decrease of 0–25% of TK mRNA in thiamine deficiency. Northern blots indicated a similar level of TK mRNA in whole brain in thiamine deficiency. These results show that the decline of TK activity results from a proportional decrease of TK protein, and the deficiency may be due to an instability of TK protein or an inhibition of TK mRNA translation. The lack of correlation of the distribution, and the absence of specific alteration, of TK in affected regions suggest that the reduced TK may not be linked directly to selective vulnerability in thiamine deficiency.     

Effectors of metabolic depression in an estivating pulmonate snail (Helix aspersa): whole animal and in vitro tissue studies

 We have examined metabolic depression in the land snail (Helix aspersa) during estivation, and have developed a tissue model of metabolic depression using an in vitro mantle preparation. The metabolic rate of H. aspersa is depressed by 84% in vivo within 4 weeks of onset of estivation, and this metabolic depression is accompanied by a decrease in haemolymph PO₂ and pH, and an increase in haemolymph PCO₂. The in vitro mantle preparation has a stable O₂ consumption and energy charge, and an energy charge similar to that of mantle in vivo. The in vitro mantle is an O₂-conforming tissue, with VO₂ varying curvilinearly with PO₂. Consequently, we have developed a mathematical method of calculating tissue VO₂ at any PO₂. These calculations show that under appropriate incubation conditions of pH and PO₂, the mantle from estivating animals shows a stable in vitro metabolic depression of 48% compared to mantle from control snails. The extrinsic effects of PO₂ and pH account for 70% of the total in vitro metabolic depression of mantle tissue; intrinsic effectors contribute a further 30%. Accepted April 26, 1996     

Differences between observed and predicted energy costs at rest and during exercise in three subsistence-level populations

Estimates of daily energy expenditure are important for many areas of research in human ecology and adaptability. The most common technique for estimating human energy expenditure under field conditions, the factorial method, generally relies on activity-specific energy costs derived from published sources, based largely on North American and European subjects. There is concern that such data may not be appropriate for non-Western populations because of differences in metabolic costs. The present study addresses this concern by comparing measured vs. predicted energy costs at rest and during sub-maximal exercise in 83 subjects (52 males, 31 females) from three subsistence-level populations (Siberian herders and high-land and coastal Ecuadorian farmers). Energy costs at rest (i.e., lying, sitting and standing) and while performing a standard stepping exercise did not significantly differ among the three groups. However, resting energy costs were significantly elevated over predicted levels (+16% in men, +11% in women), whereas exercising costs were comparable to predicted values (?6% in men, +3% in women). Elevations in resting energy needs appear to reflect responses to thermal stress. These results indicate that temperature adjustments of resting energy costs are critical for accurately predicting daily energy needs among traditionally living populations. o 1996 Wiley-Liss, Inc.     

Fasting endurance and cold resistance without hypothermia in a small predatory bird: the metabolic strategy of Tengmalm's owl,Aegolius funereus

The energetic adaptations of non-breeding Tengmalm's owls (Aegolius funereus) to temperature and fasting were studied during the birds' autumnal irruptions in western Finland. Allometric analysis (including literature data and two larger owl species measured in this study) indicates that the basal metabolic rate of owls is below the mean level of non-passerine birds. However, the basal metabolic rate of the 130-g Tengmalm's owl (1.13 W) is higher than in other owls of similar size. This is probably related to its northern distribution and nomadic life history. Relative to its size, Tengmalm's owl has excellent cold resistance due to effective insulation (lower critical temperature +10°C, minimum conductance 0.19 mW·cm^-2·°C^-1). Radiotelemetric measurements of body temperature showed that the level of body temperature is lower than for birds in general (39.4°C at zero activity) and that the amplitude of the diurnal cycle is also low (0.2–0.6°C). In contrast to many other small birds, Tengmalm's owls do not enter hypothermia during a 5-day fast at thermoneutrality or in cold. Moreover, while the metabolic rate per bird shows the expected mass-dependent decrease, the mass-specific rate decreases only slightly during the fast. In line with this, there was no decrease in the plasma triiodothyronine concentration during the fast in the owl, whereas a dramtic drop was observed in the pigeon and Japanese quail that were used as a reference. Despite this, the owl has an excellent capacity for fasting because of its ability to accumulate extensive fat depots and its low overall metabolic rate. Fasting reduced evaporative water loss to 50% of that in the fed state. Calculations show that the oxygen consumption observed in fasting birds would involve a production of metabolic water barely sufficient to compensate for evaporative water loss. The threat of dehydration may thus set a limit to the decrease in metabolic rate in fasting owls (owls rely totally on water either ingested with food or produced metabolically). We conclude that the metabolic strategy in Tengmalm's owl is largely dictated by an evolutionary pressure for fasting endurance. With the restrictions set by small body size and water economy, this bird has apparently taken these adaptations to an extreme. The constraints that preclude hypothermia, which could increase the capacity for fasting even more, remain unknown.Abbreviations BM body mass - BMR basal metabolic rate - EWL vaporative water loss - MR metabolic rate - T₃ triiodothyronine - T _a ambient temperature - T _b body temperature - VO₂ oxygen consumption     

The Western Rock lobster Panulirus cygnus (George, 1962) (Decapoda: Palinuridae): the effect of temperature and developmental stage on energy requirements of pueruli

Pueruli of the Western Rock Lobster Panulirus cygnus (George, 1962) are thought to be nonfeeding. Consequently, the metabolic rate is expected to be low during this stage in order to conserve energy reserves. Furthermore, since water temperature potentially has a substantial impact on energetic needs, the puerulus possibly exhibits mechanisms to reduce the effect of temperature on energy consumption. To test these propositions the metabolic rate was measured in post-settlement pueruli and in juveniles at two water temperatures. A respirometer of variable volume (10–50 ml) was designed for this purpose, incorporating a dark-type oxygen sensor. Results were compared with data available from the literature.

Oxygen consumption in pre-molt pueruli and in intermolt juveniles (1.48 to 5.65 μmol O₂· individual⁻¹·h⁻¹ on average) was substantially higher than in post-settlement pueruli (1.06–1.41μmol O₂·ind.⁻¹h⁻¹). These significant changes could only partly be explained through changes in biomass. Furthermore, no significant effect of an increased water temperature (from 18 to 23 ° C) could be detected on the metabolic rate in post-settlement pueruli, and the effect is moderate in pre-molt pueruli (Q₁₀ = 1.95). The water temperature has, however, a substantially greater impact on first and second molt juveniles (Q₁₀ = 2.46 to 4.80).

The energetic demand was calculated from oxygen consumption and indicate that energetic needs of post-settlement pueruli is low compared with both pre-molt pueruli and juveniles. A low energetic demand and a reduced effect of temperature on energy consumption is of considerable benefit to a non-feeding larva, and may provide the puerulus with a means of extending the duration of the non-feeding stage and increasing the chance for survival beyond metamorphosis to the first feeding stage. Results indicate that the energetic demand during metamorphosis may be considerable. It is postulated that energetic requirements of the planktonic (actively swimming) puerulus larvae are considerably higher and are likely to be more temperature dependent.     

The S11 and S13 self incompatibility alleles in Solanum chacoense Bitt. are remarkably similar

A genomic clone of the S11 allele from the self-incompatibility locus (S locus) in Solanum chacoense Bitt. has been isolated by cross-hybridization to the S. chacoense S13 allele and sequenced. The sequence of the S11 allele contains all the features expected for S genes of the Solanaceae, and S11 expression, as assessed by northern blots and RNA-PCR, was similar to that of other S. chacoense S alleles. The S11 protein sequence shares 95% identity with the phenotypically distinct S13 protein of S. chacoense and is the gametophytic S allele with the highest similarity to an existing allele so far discovered. Only 10 amino acid changes differentiate the mature proteins from these two alleles, which sets a new lower limit to the number of changes that can produce an altered S allele specificity. The amino acid substitutions are not clustered, suggesting that an accumulation of random point mutations can generate S allele diversity. The S11 intron is unusual in that it could be translated in frame with the coding sequence, thus suggesting an additional mechanism for the generation of new S alleles.     

藻－菌生态系统代谢功能的生态学研究

在室内模拟条件下,研究了一些生态因子对藻－菌（Ａ＋Ｂ）生态系统代谢有机碳（Ｃ６Ｈ１２Ｏ６）、ＮＨ３－Ｎ和无机磷（ＩＰ）的影响．研究结果表明,当藻－菌生态系统中藻（Ａ）或菌（Ｂ）的起始数量一定时,其代谢Ｃ６Ｈ１２Ｏ６的速率,随与之组合的Ｂ或Ａ的起始数量增加（数量比则相应降低）而增加．在光照和黑暗条件下,Ａ＋Ｂ系统代谢上述３种营养物质的速率均有一定的差异．黑暗下Ｃ６Ｈ１２Ｏ６的平均代谢速率较光照下高１２．３％（Ｐ＜０．０５）,ＩＰ和ＮＨ３－Ｎ的平均代谢速率则分别较光照下低１４．４％（Ｐ＜０．０５）和１６．２％（Ｐ＜０．００１）．在Ａ＋Ｂ系统和Ａ、Ｂ单培养物中,３种营养物质的代谢速率均随有机负荷量增加而增加,而且Ａ＋Ｂ系统的代谢速率分别高于单培养的Ａ和Ｂ,其中ＮＨ３－Ｎ代谢尤为显著．文章还就生态系统结构与功能的关系问题进行了讨论．