Showing papers on "ATP citrate lyase published in 2003"

Evidence of Chemolithoautotrophy in the Bacterial Community Associated with Alvinella pompejana, a Hydrothermal Vent Polychaete

Abstract: The deep-sea polychaete Alvinella pompejana colonizes tubes on the sides of black smoker chimneys along the East Pacific Rise. A diverse, yet phylogenetically constrained episymbiotic community is obligately associated with its dorsal surface. The morphologically and phylogenetically distinct dominant episymbionts have not yet been cultured, and there are no clearly defined roles for these bacteria in this symbiosis. A large insert fosmid library was screened for the presence of the two dominant phylotypes. Two fosmids, 35.2 and 38 kb, containing phylotype-specific 16S ribosmal DNA sequences were fully sequenced. Each fosmid had a gene encoding ATP citrate lyase, a key enzyme in the reverse tricarboxylic acid (rTCA) cycle, a CO2 fixation pathway. A selection of episymbiont communities from various geographic locations and vent sites were screened for the presence, diversity, and expression (via reverse transcription-PCR) of the ATP citrate lyase gene. Our results indicate that the ATP citrate lyase gene is not only a consistent presence in these episymbiont communities but is also expressed. Phylogenetically distinct forms of ATP citrate lyase were also found associated with and expressed by bacteria extracted from the tubes of A. pompejana. Utilizing PCR with degenerate primers based on a second key enzyme in the rTCA cycle, 2-oxoglutarate:acceptor oxidoreductase, we also demonstrated the persistent presence and expression of this gene in the episymbiont community. Our results suggest that members of both the episymbiont and the surrounding free-living communities display a chemolithoautotrophic form of growth and therefore contribute fixed carbon to other organisms in the vent community.

Phosphorus deficiency-induced modifications in citrate catabolism and in cytosolic pH as related to citrate exudation in cluster roots of white lupin

Abstract: A possible contribution of alterations in metabolic sequences involved in citrate catabolism, to intracellular accumulation and subsequent release of citrate was investigated in cluster roots of phosphorus (P)-deficient white lupin (Lupinus albus L.). Citrate accumulation during maturation of root clusters was associated with decreased levels of intracellular soluble Pi and ATP, and with reduced rates of respiration. Inhibitor studies with KCN and salicylhydroxamic acid (SHAM) suggest a reduced capacity of both the cytochrome pathway and of the alternative respiration with a concomitant decrease of immunochemically detectable protein levels of the alternative oxidase. Reduced respiration seems to be related to a general impairment of the respiratory system, rather than to limitation of respiratory substrates such as Pi and adenylates, as indicated by the absence of stimulatory effects of the uncoupler CCCP. The citrate/malate ratio in juvenile root clusters with high rates of respiration and low inherent levels of citrate accumulation was increased by short-term application (4–8 h) of azide and SHAM as respiration inhibitors. During maturation of root clusters, a shift from intracellular malic acid to citric acid accumulation was associated also with down-regulation of ATP citrate lyase (ACL), which catalyzes cleavage of citrate into acetyl-CoA and oxaloacetate with a putative function as anapleurotic source for the production of acetyl-CoA under P-deficient conditions. Inhibition of nitrate uptake and assimilation is a general response to P limitation in many plant species including white lupin. Reduced consumption of the amino acceptor 2-oxoglutaric acid as a product of citrate turnover may therefore contribute to increased citrate accumulation. Accordingly, artificial inhibition of nitrate reduction by localized application of tungstate significantly increased the citrate/malate ratio in juvenile root clusters. Lowering the cytosolic pH by external application of propionate stimulated citrate and malate exudation in non-cluster lateral roots and in developing root clusters. This effect was reverted by preincubation with phosphonate to buffer the cytosol. The results suggest that acidification of the cytosol may be an important factor, triggering the transient release of citrate and protons from mature root clusters in P-deficient white lupin.

Effects of altered citrate synthase and isocitrate dehydrogenase expression on internal citrate concentrations and citrate efflux from tobacco (Nicotiana tabacum L.) roots

Abstract: To assess the effectiveness of manipulating citrate metabolism with the aim of increasing citrate efflux from roots, we generated transgenic tobacco (Nicotiana tabacum L.) lines that either overexpressed mitochondrial citrate synthase (EC 4.1.3.7) activity or had reduced activity of cytosolic isocitrate dehydrogenase (EC 1.1.1.42). Despite increases in citrate synthase activities in transgenic lines of up to 5-fold, neither internal citrate concentrations nor citrate efflux were increased compared to controls suggesting that, in tobacco, citrate synthase activity does not directly determine citrate accumulation and efflux. Consistent with a lack of effect on citrate efflux, the increase in citrate synthase activity did not enhance the aluminium resistance of the transgenic lines. Preliminary data collected on two transgenic lines with cytosolic isocitrate dehydrogenase activities reduced to one-tenth and one third of the control for shoot and root tissues respectively, showed that while these changes in activities were associated with a 1.5-fold increase in internal citrate concentrations of both types of tissue, citrate efflux from roots was not increased. Further work is needed to establish whether the increase in internal citrate concentration is associated with enhanced aluminium resistance of these lines. We conclude that in tobacco internal citrate concentrations and citrate efflux are largely insensitive to large changes in either mitochondrial citrate synthase or cytosolic isocitrate dehydrogenase activities and suggest that other factors, such as transport out of the roots, control citrate efflux.

Metabolic function of hepatic hexokinase in perch, Perca fluviatilis

Abstract: Feeding previously fasted perch with a carbohydrate-rich pelleted diet resulted in a rapid transient insulin response followed by a transient increase in the activity of the hepatic low-affinity hexokinase (glucokinase, GK). The activity of the high-affinity hexokinase (hexokinase, HK) increased more slowly and steadily to about seven times the fasting value on the 10th day after start of feeding, in parallel with an increase in the hepatic glycogen content. An insulin response was also observed when fasted perch were fed a diet consisting of boiled fillet of cod (a high-protein, very low-fat and practically carbohydrate-free diet). In this case no GK activity was observed, whereas the glycogen content increased steadily from the start of feeding to about the same level as when fish were given the carbohydrate-rich diet. There was a delayed increase in the activity of HK, starting on about the sixth day (after the glycogen stores were filled), concomitant with increased activities of lipogenic enzymes (glucose-6-phosphate dehydrogenase, ATP citrate lyase and malic enzyme). The results are in line with the hypothesis that the mitochondria-bound hepatic HK has a particular anabolic function in glycogen and fat synthesis.

Activities of Enzymes Involved in Fatty Acid Metabolism in the Colon Epithelium of Piglets Fed with Different Fiber Contents Diets

Abstract: The present study was conducted to evaluate the influence of dietary fiber on the activities of malic enzyme and citrate lyase involved in fatty acid metabolism in the colon epithelium of pigs. Thirty-six weaned 5 weeks old crossbred (Yorkshire×Swedish Landrace) piglets originating from twelve litters were randomly assigned to either a low fiber diet containing 10% non-starch polysaccharides (NSP), a control diet containing 14.7% NSP or a high fiber diet containing 20% NSP. The activity of malic enzyme in the colonic epithelium of pigs significantly (p<0.05) increased with age during the suckling-weaning transition. There was a tendency (p<0.10) of decreased malic enzyme activity in the colonic epithelium of pigs fed on the high fiber diet. At week 6, a lowered (p<0.01) activity of malic enzyme in pigs fed on the low fiber diet compared with that in pigs fed on the high fiber and the control diets. Nevertheless, there were no significant differences in the activity of citrate lyase observed either between pigs with different ages or between pigs fed with various diets. The current data suggest that piglets during the suckling-weaning transition have a limited capacity to synthesize fatty acids from carbohydrate derivatives in the coloncytes. In addition, lipogenesis in coloncytes was enhanced with age during the suckling-weaning transition. A tendency (p<0.10) to an increased capacity to utilize acetyl-CoA in coloncytes of pigs has been observed for the high fiber diet. Moreover, the present work indicated that dietary fiber resulted in a lowered rate of lipogenesis and a reduced activity of malic enzyme. (Asian-Aust. J. Anim. Sci. 2003. Vol 16, No. 10 : 1524-1528)

Medicinal composition for inhibiting the expression of atp-citrate lyase and use thereof

Abstract: It is intended to provide a medicinal composition for inhibiting the expression of ATP-citrate lyase which contains a compound inhibiting the expression of ATP-citrate lyase when administered into the living body. Examples of the compound inhibiting the expression of ATP-citrate lyase when administered into the living body include insulin secretion accelerators such as nateglinide. This medicinal composition is efficacious in preventing, ameliorating and treating metabolic syndromes, in particular, liver disorders relative to lipid metabolic errors such as fatty liver and NASH.