Global gene expression of Deinococcus radiodurans, a highly radiation resistant bacterium, in response to excess copper was analyzed by using oligonucleotide microarray chip. Among 3,187 open reading frames of D. radiodurans, seventy genes showed a statistically significant expression ratio of at least 2-fold changes under growth conditions of excess copper; 64 genes were induced and 6 genes were reduced. Especially, two operons ($DRB0014{\sim}DRB0017$ and $DRB0125{\sim}DRB0121$) presumably involved in the iron transport and utilization were the most highly induced genes by excess copper. A quantitative real-time PCR assay revealed that DRB00l4 and DRB0125 are highly transcribed responding to excess copper and 2,2'-dipyridyl, an iron chelator. In addition, the transcription of both genes was not changed by excess iron and bathocuproine disulphonate, a copper chelator. These results suggested that the copper metabolism may be closely connected with the iron transport and utilization in D. radiodurans. However, the disruption of each gene, DRB00l4 and DRB0125, did not affect the copper and radiation resistance, the most well-known character of this organism.
Roy, Swapan Kumar;Kwon, Soo Jeong;Cho, Seong-Woo;Kamal, Abu Hena Mostafa;Kim, Sang-Woo;Sarker, Kabita;Jeong, Hae-Ryong;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.128-128
/
2017
Copper (Cu) is very toxic to plant cells due to its inhibitory effects on many physiological and biochemical processes. In spite of its potential physiological and economic significance, molecular characterization after Cu stress has so far been grossly overlooked in sorghum. To explore the molecular alterations that occur in response to copper stress, the present study was executed in ten-day-old Cu-exposed leaves of sorghum seedlings. The growth of shoots was markedly reduced, and ionic alterations were prominently observed in the leaves when the seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CuSO_4$. Using two-dimensional gels with silver staining, 643 differentially expressed protein spots (${\geq}1.5-fold$) were identified as either significantly increased or reduced in abundance. Of these spots, a total of 24 protein spots (${\geq}1.5-fold$) from Cu-exposed sorghum leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Of the 24 differentially expressed proteins from Cu-exposed sorghum leaves, a total of 13 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of most identified protein species, which function in carbohydrate metabolism, stress defense, and protein translation, was significantly enhanced, while that of another protein species involved in energy metabolism, photosynthesis and growth and development were severely reduced. The resulting differences in protein expression patterns together with related morpho-physiological processes suggested that these results could help to elucidate plant adaptation to Cu stress and provide insights into the molecular mechanisms of Cu responses in $C_4$ plants. The over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu.
Heavy metals which are present as trace elements in human body have been known to modify various enzymatic reaction. These metals can be essential or non-essential. Zinc, copper and calcium are essential in maintaining some biological processes, whereas non-essential metals such as cadmium, lead and mercury produce accumulatve toxic effect. Cadmium accumulated in pancreas can cause toxicity and damage of pancreatic cells, thereby influencing CHO metabolism. Lead compounds are known to produce toxic effects on the kidney, digestive system and brain fellowed by inhibition of activity of ${\rho}-aminolevulinic$ acid and biosynthesis of hemoproteins and cytochrome. Evidence has been accumulated that zinc not only acts as a cofactor in enzyme reaction but also prevents toxic effect induced by heavy metal such as copper and cadmium. To demonstrate the effect of heavy metals on pancreatic secretion, part of uncinate pancreas was taken and incubated in Krebs-Ringer bicarbonate buffer with heavy metals used. Additional treatment with CCK-OP was performed when needed. After incubation during different period of time, medium was analyzed for amylase activity using Bernfeld's method. The present study was attempted in order to elucidate the effect of several kinds of heavy metal on exocrine pancreatic secretion in vitro. The results obtained are as follows: 1) CCK-OP stimulated significantly amylase release from pancreatic fragments in vitro. 2) CCK-OP response of amylase release from pancreatic fragments was inhibited by treatmant with cadmium, especially high doses of cadmium. 3) CCK-OP response of amylase release from pancreatic fragments was inhibited when pretreated with $10^{-4}M$ copper chloride. 4) Lead chloride at the concentration of $10^{-3}M\;and\;10^{4}M$ stimulated the basal amylase release in vitro but CCK-OP response did not augment by lead chloride. 5) Zine chloride did not affect amylase release from pancreatic fragment in vitro. From the results mentioned above, it is suggested that CCK-OP response was inhibited it the amylase release from pancreatic fragments pretreated with cadmium and copper chloride.
Plants need essential mineral elements to favorably develop and to complete their life cycle. Despite the irreplaceable roles of microelements, they are often ignored due to the relative importance of macroelements with their influence on crop growth and development. We focused on the changes in primary metabolites in the leaves and roots of bell pepper plants under 6 microelements-deficient conditions: Copper (Cu), Zinc (Zn), Iron (Fe), Manganese (Mn), Boron (B) and Molybdenum (Mo). Bell pepper plants were grown in hydroponic containers, and individual elements were adjusted to 1/10-strength of Hoagland nutrient solution. A remarkable perturbation in the abundance of the primary metabolites was observed for the Fe and B and the Mn and B deficiencies in the leaves and roots, respectively. The metabolites with up-accumulation in the Fe-deficient leaves were glucose, fructose, xylose, glutamine, asparagine and serine. In contrast, the Mn deficiency also resulted in a higher accumulation of glucose, fructose, xylose, galactose, serine, glycine, β-alanine, alanine and valine in the roots. The B deficiency noticeably accumulated alanine, valine and phenylalanine in the roots while it showed a substantial decrease in glucose, fructose and xylose. These results show that the primary metabolism could be seriously disturbed due to a microelement deficiency, and the alteration may be either the specific or adaptive responses of bell pepper plants.
Twenty eight 3-4 month old castrated Black Bengal kids (Capra hircus) were used to determine the effects of source and level of dietary copper (Cu) concentration on their performance and nutrient utilization. Cu was supplemented (0, 10, 20 and 30 mg/kg diet DM) as copper sulfate ($CuSO_4$, $5H_2O$) or copper proteinate (Cu-P). Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each) and mineral and vitamin mixture (0.5% each) supplements at 3.5% of body weight to meet NRC (1981) requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. $CuSO_4$ or Cu-P was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well ventilated shed with facilities for individual feeding in aluminum plated metabolic cages. Blood samples were collected from the jugular vein on d 0, 30, 60 and 90 to determine hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and serum enzymes (alkaline phosphatase, alanine transferase and aspertate transferase). A metabolism trial of 6 days duration was conducted after 90 days of experimental feeding. Statistical analysis revealed that source and level of Cu supplementation improved live weight gain (p<0.04) and average daily gain (p<0.01). No significant contribution of source and level of Cu to alter serum serum enzymes was evident. Goats fed Cu-P tended to have higher Hb, PCV and TEC than with $CuSO_4$ supplementation. Cu-P increased digestibility of ether extract (EE, p<0.02) and crude fiber (p<0.05) and showed an increasing trend (p<0.09) for digested crude protein (CP) and crude fiber (CF). Supplemental dose of Cu linearly improved (p<0.02) digestibilities of dry matter (DM), organic matter (OM), EE and nitrogen free extract (NFE). Though the absorption of nitrogen (N) was not affected (p>0.10) by both source and dose of Cu, N retention was affected (p<0.04) and there was a significant $Source{\times}Dose$ interaction (p<0.05). Final body weight (BW) was not influenced (p>0.10) by the source of Cu but increasing dose of Cu increased (p<0.04) the BW of kids. TDN intake (g/kg $W^{0.75}$) was higher (p<0.05) with the increased dose of Cu and there was a significant $Source{\times}Dose$ interaction. It was concluded that supplementation of Cu from different sources and varying dose level in a concentrate based diet may improve performance, nutrient utilization and plane of nutrition in castrated Black Bengal kids. The effects on performance and nutrient utilization are more pronounced with Cu-P than $CuSO_4$ supplementation. Higher dose of Cu showed better result than lower dose.
Chlorella ellipsoidea cells were cultured in an iron, copper, zinc, manganese, molybdenum or boron-free medium. Biosynthetic activities of nucleic acids, protein and phospholipid in chlorella cells, which were growing in a microelement deficient medium were compared with those of the normal cells by measuring the contents of phosphate, amino acids or UV-absorbing substances in the various cell fractions. When the algae were grown in a molybdenum-free medium, the amounts of phosphate in the acid-soluble fraction of the cells increased, whereas the amounts of alkali-stable protein and RNA decreased compared with the normal cells showing that the synthesis of protein and RNA from the early products of photosynthesis was inhibited. When the algae were grown in a boron-free medium, amounts of alkali-labile protein and phospholipid of the cells decreased, while the amount of phosphate in acid-soluble fraction increased compared with the normal cells showing that the biosynthesis of protein and phospholipid from the early products of photosynthesis was retarded. In general, amounts of protein and RNA in the microelement deficient cells significantly decreased compared with those of the normal cells. Phosphate content in the acid-soluble fraction of the algal cell grown in an zinc, copper, molybdenum, or boron-free medium increased considerably, whereas that of the algal cell grown in an iron or manganese-free medium decreased remarkably compared with that of the control. It is considered, therefore, that molybdenum, zinc, copper and boron etc. play an important role in the biosyntbesis of macromolecule from acid-soluble phosphate compounds, in contrast to the principal action of iron and manganese on the photosynthetic process itself.
Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.130-130
/
2017
Sorghum bicolor is considered as copper-tolerant species. The present study was conducted to understand the copper tolerance mechanism in Sorghum seedling roots. Morphological and effects of Cu on other interacting ions were observed prominently in the roots when the plants were subjected to different concentrations (0, 50, and $100{\mu}M$) of $CuSO_4$. However, the morphological characteristics were reduced by Cu stress, and the most significant growth inhibition was observed in plants treated with the highest concentration of $Cu^{2+}$ ions ($100{\mu}M$). In the proteome analysis, high-throughput two-dimensional polyacrylamide gel electrophoresis coupled with MALDI-TOF-TOF mass spectrometry was performed to explore the molecular responses of Cu-induced sorghum seedling roots. In two-dimensional silver-stained gels, a total of 422 differentially expressed proteins (${\geq}1.5-fold$) were identified using Progenesis SameSpot software. A total of 21 protein spots (${\geq}1.5-fold$) from Cu-induced sorghum roots were analyzed by mass spectrometry. Of the 21 differentially expressed protein spots from Cu-induced sorghum roots, a total of 10 proteins were up-regulated, and 11 proteins were down-regulated. The abundance of the most identified protein species from the roots that function in stress response and metabolism was significantly enhanced, while protein species involved in transcription and regulation were severely reduced. The results obtained from the present study may provide insights into the tolerance mechanism of seedling roots in Sorghum.
The primary objective of these studies was to screen the materials showing inhibitions of HMG-CoA reductase in vitro. The secondary objective was to determine the effect of garlic, lovastatin and copper on cholesterol concentrations in plasma. liver and brease muscle of pullets. In experiment 1, the degree of inhibition of the selective samples on HMG-CoA reductase activity was determined in vitro. The inhibition rate of water soluble garlic extracts, lovastatin and copper to HMG-coA reductase activity were 51.3%, 87.5%, and 82.0% respectively . In experiment 2, control diet (basal diet), garlic powder (3% in diet) , lovastatin (300mg/kg of diet) and copper(200mg/kg of diet) were fed to pullets in order to investigate the changes of cholesterol concentration in plasma and tissues. Plasma total cholesterol , and LDL-cholesterol were significantly reduced in pullets fed a diet containing 3 % garlic powder. However, coper significantly increased total cholesterol compared to controls and lovastatin did not affect plasma chholesterol concentration . Total cholesterol inlover and breast muscle inpullets were not affectedb y adding cholesterol lowering materials to the diets. The data suggests that it is not easy for HMG-CoA reductase inhibitors to reduce cholesterol levels in the body due to complication in cholestrol metabolism . However, garlic administration can lower the levels of plasma cholesterol in pullets.
Metal binding proteins or metallo-proteins are important for the stability of the protein and also serve as co-factors in various functions like controlling metabolism, regulating signal transport, and metal homeostasis. In structural genomics, prediction of metal binding proteins help in the selection of suitable growth medium for overexpression's studies and also help in obtaining the functional protein. Computational prediction using machine learning approach has been widely used in various fields of bioinformatics based on the fact all the information contains in amino acid sequence. In this study, random forest machine learning prediction systems were deployed with simplified amino acid for prediction of individual major metal ion binding sites like copper, calcium, cobalt, iron, magnesium, manganese, nickel, and zinc.
Ceruloplasmin (Cp) is a copper protein with important functions in iron homeostasis and in inflammation. Cp mRNA expression is induced by interferon (IFN)-${\gamma}$ in U937 monocytic cells, but synthesis of Cp protein is halted after about 12 h by transcript-specific translational silencing. The silencing mechanism requires binding of a 4-component cytosolic inhibitor complex, IFN-gamma-activated inhibitor of translation (GAIT), to a defined structural element (GAIT element) in the Cp 3'-UTR. Translational silencing of Cp mRNA requires the essential proteins of mRNA circularization, suggesting that the translational inhibition requires end-to-end mRNA closure. These studies describe a new mechanism of translational control, and may shed light on the role that macrophage-derived Cp plays at the intersection of iron homeostasis and inflammation.
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