Rise In Protein Synthesis Research Articles

Impacts of Wheat Powdery Mildew on Grain Yield & Quality and Its Prevention and Control Methods

As one of the major foliar diseases, wheat powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) affects photosynthesis of nutrient organs, such as stems and leaves, causing a reduction in production to varying degrees. In recent years, it has been found that powdery mildew can also affect grain quality, including nutritional quality and processing quality. Powdery mildew can cause changes of grain protein content， its components, starch content and ratio of amylose to amylopectin, and it can also cause changes of grain volume weight, flour falling number and viscosity, extensograph and farinograph parameters, etc. The studies have also found that powdery mildew can increase the expression level of molecular chaperones and enzymes relating to protein synthesis, which led to the rise in protein synthesis. And glycolysis-related proteins also show high expression levels in grains from infected wheat, which implies that glycolysis is very active and further results in the depletion of starch content. These results show that the research on the mechanism of quality change caused by disease has also made some progress.This paper includes five parts: the first part introduces the infection characteristics of wheat powdery mildew. The second part elaborates the change of wheat yield caused by powdery mildew and its influence mechanism. In the third part, the effect of powdery mildew on wheat quality and its molecular mechanism are discussed. The fourth part analyzes the four prevention and control ways from cultivating disease-resistant varieties, strictly controlling the use of fungicides, advocating spraying silicate and biological control. Finally, the last part is a prospect where the effect of wheat powdery mildew on quality and its molecular mechanism and the comprehensive pollution-free control system of wheat powdery mildew are discussed. The paper aims to provide the basis for the research of quality physiology and pollution-free control of wheat powdery mildew.

Divergent Anabolic Signalling responses of Murine Soleus and Tibialis Anterior Muscles to Chronic 2G Hypergravity

The purpose of the study was to assess the rate of protein synthesis (PS) and elucidate signalling pathways regulating PS in mouse soleus (Sol) and tibialis anterior (TA) muscles following chronic hypergravity (30-day centrifugation at 2G). The content of the key signalling proteins of the various anabolic signalling pathways was determined by Western-blotting. The rate of PS was assessed using in-vivo SUnSET technique. An exposure to 2G centrifugation did not induce any significant changes in the rate of PS as well as phosphorylation status of the key anabolic markers (AKT, p70s6k, 4E-BP1, GSK-3beta, eEF2) in Sol. On the contrary, a significant 55% increase in PS (p < 0.05) was found in TA. The cause of such a rise in PS could be associated with an increase in AKT (+72%, p < 0.05), GSK-3beta (+60%, p < 0.05) and p70s6k (+40%, p < 0.05) phosphorylation, as well as a decrease in eEF2 phosphorylation (−46%, p < 0.05) as compared to control values. Thus, the results of our study indicate that 30-day 2G centrifugation induces a distinct anabolic response in mouse Sol and TA muscles. The activation of the PS rate in TA could be linked to an up-regulation of both mTORC1-dependent and mTORC1-independent signalling pathways.

Model of the delayed translation of cyclin B maternal mRNA after sea urchin fertilization.

Sea urchin eggs exhibit a cap-dependent increase in protein synthesis within minutes after fertilization. This rise in protein synthesis occurs at a constant rate for a great number of proteins translated from the different available mRNAs. Surprisingly, we found that cyclin B, a major cell-cycle regulator, follows a synthesis pattern that is distinct from the global protein population, so we developed a mathematical model to analyze this dissimilarity in biosynthesis kinetic patterns. The model includes two pathways for cyclin B mRNA entry into the translational machinery: one from immediately available mRNA (mRNAcyclinB) and one from mRNA activated solely after fertilization (XXmRNAcyclinB). Two coefficients, α and β, were added to fit the measured scales of global protein and cyclin B synthesis, respectively. The model was simplified to identify the synthesis parameters and to allow its simulation. The calculated parameters for activation of the specific cyclin B synthesis pathway after fertilization included a kinetic constant (ka ) of 0.024 sec-1 , for the activation of XXmRNAcyclinB, and a critical time interval (t2 ) of 42 min. The proportion of XXmRNAcyclinB form was also calculated to be largely dominant over the mRNAcyclinB form. Regulation of cyclin B biosynthesis is an example of a select protein whose translation is controlled by pathways that are distinct from housekeeping proteins, even though both involve the same cap-dependent initiation pathway. Therefore, this model should help provide insight to the signaling utilized for the biosynthesis of cyclin B and other select proteins. Mol. Reprod. Dev. 83: 1070-1082, 2016. © 2016 Wiley Periodicals, Inc.

The effect and molecular mechanism of powdery mildew on wheat grain prolamins

SUMMARYA field experiment was conducted to investigate the effects of powdery mildew (Blumeria graminis f. sp. Tritici, Bgt) on wheat grain at varying levels of disease severity and at different growth stages. Methods used to determine these effects included Kjeldahl determination, unidimensional polyacrylamide gel electrophoresis, dielectrophoresis combined with mass spectrometric analysis. The specific influences explored were those on prolamins and protein composition at the molecular level. Concentrations of both grain protein and prolamin in wheat increased as disease indices (DIs) of powdery mildew rose from 20 days after anthesis (DAA) to maturity. Globulin concentrations changed dynamically and significantly, especially at 25 DAA when DI was the highest. This was verified by proteomic analysis which showed globulins (such as globulin 3, globulin 3B, globulin 3C, gliadin/avenin-like protein and triticin) being up-regulated significantly under powdery mildew stress. It was proposed that powdery mildew might indirectly affect protein accumulation in grain by influencing the regulative enzymes (including peptidylprolyl isomerase, cyclophilin A-2 and GTPase ObgE) and metabolic processes. It was speculated that the indirect increase caused by yield reduction was not the only factor causing the increase in prolamin concentration. Another factor may be the rise of expression level of molecular chaperones and enzymes relating to protein synthesis, which led to the rise in protein synthesis.

EIF4E/4E-BP dissociation and 4E-BP degradation in the first mitotic division of the sea urchin embryo

The mRNA’s cap-binding protein eukaryotic translation initiation factor (eIF)4E is a major target for the regulation of translation initiation. eIF4E activity is controlled by a family of translation inhibitors, the eIF4E-binding proteins (4E-BPs). We have previously shown that a rapid dissociation of 4E-BP from eIF4E is related with the dramatic rise in protein synthesis that occurs following sea urchin fertilization. Here, we demonstrate that 4E-BP is destroyed shortly following fertilization and that 4E-BP degradation is sensitive to rapamycin, suggesting that proteolysis could be a novel means of regulating 4E-BP function. We also show that eIF4E/4E-BP dissociation following fertilization is sensitive to rapamycin. Furthermore, while rapamycin modestly affects global translation rates, the drug strongly inhibits cyclin B de novo synthesis and, consequently, precludes the completion of the first mitotic cleavage. These results demonstrate that, following sea urchin fertilization, cyclin B translation, and thus the onset of mitosis, are regulated by a rapamycin-sensitive pathway. These processes are effected at least in part through eIF4E/4E-BP complex dissociation and 4E-BP degradation.

Chemotropic Responses of Retinal Growth Cones Mediated by Rapid Local Protein Synthesis and Degradation

Growth cones contain mRNAs, translation machinery, and, as we report here, protein degradation machinery. We show that isolated retinal growth cones immediately lose their ability to turn in a chemotropic gradient of netrin-1 or Sema3A when translation is inhibited. Translation inhibition also prevents Sema3A-induced collapse, while LPA-induced collapse is not affected. Inhibition of proteasome function blocks responses to netrin-1 and LPA but does not affect Sema3A responses. We further demonstrate in isolated growth cones that netrin-1 and Sema3A activate translation initiation factors and stimulate a marked rise in protein synthesis within minutes, while netrin-1 and LPA elicit similar rises in ubiquitin-protein conjugates. These results suggest that guidance molecules steer axon growth by triggering rapid local changes in protein levels in growth cones.

EIF4E Association with 4E-BP Decreases Rapidly Following Fertilization in Sea Urchin

The eukaryotic translation initiation factor (eIF) 4F facilitates the recruitment of ribosomes to the mRNA 5′ end. The 4E-BPs are small proteins with hypophosphorylated forms that interact with the cap binding protein eIF4E, preventing its interaction with eIF4G, thereby preventing ribosome interaction with mRNA. In sea urchin, fertilization triggers a rapid rise in protein synthesis. Here, we demonstrate that a 4E-BP homologue exists and is associated with eIF4E in unfertilized eggs. We also show that 4E-BP/eIF4E association diminishes a few minutes following fertilization. This decrease is correlated with a decrease in the total amount of 4E-BP in combination with an increase in the phosphorylation of the protein. We propose that 4E-BP acts as a repressor of protein synthesis in unfertilized sea urchin eggs and that 4E-BP/eIF4E dissociation plays an important role in the rise in protein synthesis that occurs shortly following fertilization.

Changes in fast axonal transport in sensory neurons during tadpole metamorphosis

Fast axonal transport of radiolabeled protein was examined in lumbar and tail dorsal root ganglion (DRG) neurons at progressive stages of bullfrog tadpole metamorphosis. Accumulation of [35S]methionine-labeled protein proximal to a lumbar peripheral nerve ligature (at a fixed distance from the DRG) increased as tadpoles advanced from premetamorphosis through prometamorphosis to metamorphic climax. The rate of increase was steeper when expressed as a percentage of protein synthesized in the neurons of origin than when expressed as a percentage of total DRG protein synthesis. Further, the increase was not secondary to a rise in protein synthesis. In contrast, fast axonal transport decreased in DRG neurons of the tail at the onset of metamorphic climax, when tail resorption is initiated. The stage-related increase in protein transport in lumbar nerves is due, at least in part, to an increased rate of transport. As determined from optically detected anterograde organelles in individual lumbar nerve axons, an approximate doubling of the fast transport rate occurred between the premetamorphic stage and metamorphic climax. In addition, the rates of organelle transport in lumbar axons of adult bullfrogs were significantly greater than in corresponding axons of tadpoles at metamorphic climax, further suggesting that organelle velocity is a developmentally regulated parameter of fast axonal transport.

Chapter 56: Progressive increases of protein synthesis in the circumventricular organs during chronic dehydration in rats

The quantitative autoradiographic method with L-(35S)methionine was applied to investigate the effect of chronic dehydration on rates of protein synthesis in circumventricular organs (CVOs). Water deprivation for 1, 2 and 3 days causes progressive increases of protein synthesis in the subfornical organ (SFO), the area postrema, the organum vasculosum laminae terminalis and the neurohypophysis. Chronic salt ingestion with 2% NaCl in drinking water for 3 days resulted in increases of protein synthesis in the CVOs similar to those found after 3 days water deprivation, with only one exception, the SFO, in which the rise in protein synthesis was of lower amplitude after 3 days salt ingestion as compared to 3 days water deprivation. These results suggest that several circulating factors related to intracellular dehydration and the high plasma levels of the neurohormones vasopressin and oxytocin are probably important determinants of the rise of protein synthesis in circumventricular organs. Alternatively, the elevated level of blood-borne angiotensin II may well explain the higher metabolic response of the SFO following water deprivation compared to salt ingestion.

A major substrate of maturation promoting factor identified as elongation factor 1 beta gamma delta in Xenopus laevis

Protein synthesis is believed to be under control of the cell cycle during meiosis and mitosis. Any relationship between substrates for cdc2 kinase and components of the protein synthetic apparatus would therefore be of prime importance. During meiosis of Xenopus laevis oocytes one of the substrates for this kinase is a p47 protein, which is complexed to two other proteins, P36 and P30. Judged from partial amino acid sequence data on P47 and P30, the P30 and P47 proteins were reported to resemble the protein synthetic elongation factors (EF) 1 beta and 1 gamma from Artemia salina (Bellé, R., Derancourt, J., Poulhe, R., Capony, J.P., Ozon, R., and Mulner-Lorillon, O. (1989) FEBS Lett. 255, 101-104). This paper shows that the complex composed of P30, P47, and P36 from Xenopus is identical to the complex of EF-1 beta, EF-1 gamma, and EF-1 delta from Artemia according to two criteria. 1) Both stimulate elongation factor 1 alpha-mediated transfer RNA binding to ribosomes and exchange of guanine nucleotides on elongation factor 1 alpha to a comparable degree. 2) Each of the three subunits of the protein complex P30.P47.P36 from Xenopus shows a structural homology with one of the corresponding subunits of EF-1 beta gamma delta from Artemia. Presumably the phosphorylation of EF-1 gamma, which associates with tubulin at least in vitro, is important in processes following the onset of meiosis which is accompanied by a rise of protein synthesis.

Does hyperketonemia affect protein or glucose kinetics in postabsorptive or traumatized man?

Leucine and glucose turnover were measured using simultaneous infusions of [ 13C]leucine and [ 2H]glucose before and during an infusion of Na dl-hydroxybutyrate (Na dl-HB) in overnight-fasted patients the day before and 3 days after total hip replacement. The ketone body infusion before surgery resulted in a significant increase in plasma leucine concentration and leucine turnover, while glucose concentration and turnover decreased. Surgery increased leucine turnover. Ketone body infusion after surgery caused a further increased leucine turnover while turnover fell as before surgery. We suggest that exogenous ketone bodies decrease hepatic glucose production and probably stimulate a rise in protein synthesis above breakdown leading to a decreased nitrogen excretion as observed by other investigators. Despite the metabolic adaptation to trauma, this response was not affected by surgery.

The Effect of Refeeding on Tissue Protein Synthesis in Rainbow Trout

The fractional rate of protein synthesis ($K_{s}$ %/day-i.e., the percentage of tissue protein synthesized per day) was measured in various tissues of 6-day fasted rainbow trout at 3, 6, and 12 h after being refed. There was a significant stimulation in the rate of protein synthesis in all of the tissues except the white muscle by 3 h after refeeding. The red muscle, white muscle, and ventricle exhibited a slow, graded response, with the white muscle not reaching levels found in continually fed fish 12 h after the meal. The gill, stomach, and intestine showed a rapid rise in protein synthesis reaching a plateau after approximately 3 h. The liver exhibited a peak in protein synthesis within 3 h that subsequently declined. Generally, there was no effect of refeeding on tissue RNA/protein ratio (used to indicate tissue ribosome concentration) so that these changes in tissue protein synthesis were brought about through an increase in protein synthesis per unit RNA (which is indicative of an alteration in the rate of ribosomal translation). Following the injection of the prostaglandin synthesis inhibitor, indomethacin, only in the gill was the refeeding stimulation partially blocked, implicating the prostaglandins in the regulation of protein synthesis in this tissue.

Escape from X-Ray-Induced Arrest for Lens Cells Stimulated from Quiescence: Time Relationship to RNA, Protein, and DNA Synthesis

Quiescent cells of the central zone region of the rat lens epithelium were stimulated to enter the proliferation cycle by wounding. RNA synthesis and a corresponding increase in poly(A)+/total RNA reached a peak by Hour 4. Cells progressed into the G1B compartment by Hour 10. A rise in protein synthesis began at Hour 8, and onset of DNA synthesis occurred by Hour 14. The timing of cell cycle progression that allowed escape from a dose of X irradiation that completely inhibited DNA synthesis was investigated. A growth-arrest point was identified at Hour 9 where 10 GY of X irradiation given before, but not after, completely inhibited earliest responding cells from entering DNA synthesis on schedule. Increased quantities of cells entered DNA synthesis on schedule as timing of the X irradiation was moved closer to the end of G1. Based on time relationships, the rise in protein synthesis is correlated with the "sufficient" event for the escape.

Ectromelia virus-induced changes in primary cultures of mouse hepatocytes.

Mouse hepatocytes were isolated by collagenase perfusion, maintained in non-proliferating monolayer culture and shown to retain liver cell function as judged by gluconeogenesis for 15 to 18 h. Such cells could be infected with and support the replication of a virulent strain of ectromelia virus. Virus antigen and characteristic cytoplasmic 'B'-type poxvirus inclusion bodies were demonstrated by immunofluorescence in virtually all cells. By electron microscopy it was shown that 'B'-type inclusions were the site of virus replication, and that the biogenesis of ectromelia virus and ultrastructural changes in hepatocytes were similar to those observed in infected mouse livers. Early cell rounding effects, a normal characteristic of poxvirus infections in tissue culture cells, were not seen in ectromelia-infected hepatocytes, although late degenerative changes did occur. Pulse-labelling of hepatocyte cultures with [35S]methionine showed that ectromelia virus inhibited the rise in protein synthesis seen in controls and imposed a gradual decline in host protein synthesis to an extent and at a rate significantly different from that in mouse L929 cells. Gluconeogenesis was inhibited by ectromelia virus infection of hepatocytes.

The effect of indomethacin on the stimulation of protein synthesis by insulin in young post-absorptive rats.

Groups of young rats (100 g body wt.) were starved from 23:00 to 11:00 h. The animals were then infused intravenously with diluent or insulin at three different doses to achieve plasma insulin concentrations of 20, 50 and 150 microunits/ml. Before the start of the infusion, animals received a single intravenous injection of indomethacin (250 micrograms) or diluent. After 20 min of infusion, the rats were injected with a large amount of labelled phenylalanine and were killed 10 min later. Insulin produced a dose-dependent decrease in plasma glucose and a dose-dependent rise in protein synthesis in cardiac, gastrocnemius, plantaris and soleus muscles. Protein synthesis in the liver was unaffected by insulin. Indomethacin had no effect on plasma glucose concentrations, but blocked the insulin-induced rise in protein synthesis in cardiac, gastrocnemius and plantaris, but not in soleus muscle. The hormone also increased the plasma concentration of prostaglandin E2 and of prostaglandins F2 alpha and E2 in gastrocnemius and plantaris muscle. The results show close similarities to previous observations with isolated rabbit muscles in vitro and suggest that the involvement of arachidonic acid metabolism in the action of insulin on protein synthesis is of physiological significance.

Polysome structure in sea urchin eggs and embryos: An electron microscopic analysis

Modified chromatin-spreading techniques have been used to prepare reproducible spreads of polysomes for EM visualization. We have analyzed polysomes of sea urchin ( Lytechinus pictus) eggs and embryos to elucidate some of the events which occur during the burst of protein synthetic activity following fertilization. We have confirmed that the rise in protein synthesis after fertilization is concomitant with the recruitment of messenger RNA molecules into polysomes. The presence of ribosome-free tails is a structural feature of polysomes in which the mRNA is being translated for the first time. This structural marker has allowed us to derive an elongation rate of 1.5–1.8 amino acids/sec and a transit time of 5 min for the average-sized polysome in L. pictus embryos at 16°C. We have also observed a difference between the structure and the average size of polysomes in the egg and embryo. Egg polysomes are longer (average size = 11.96 ribosomes, n = 671) than 1-hr embryo polysomes (average size = 7.14 ribosomes, n = 863) and 10% of the egg polysomes contain visible gaps while less than 1% of the 1-hr embryo polysomes contain internal ribosome-free regions. We speculate that such differences reflect the lower translation efficiency of the relatively dormant egg cell.

Isoenzymic changes in myosin and hypertrophy; adaptation during chronic mechanical overload.

Hypertrophy and a decrease in maximal contractile rate characterize the cardiac response to chronic mechanical overload. Both protect the myocardium, the first by increasing the number of contractile units, the second by maintaining the normal energy balance. Decrease in the contractile rate is correlated with (and is probably a function of) the decrease in myosin ATPase. This is due to an isoenzymic change, which has been demonstrated in the rat but remains hypothetical in man, towards a fetal form with a low ATPase activity. This redistribution, correlated with the degree of hypertrophy, is seen after stenosis or aortic insufficiency, infarction, a genetic tendency to hypertension and, after a longer latency, aorto-caval fistulae. The maximal contractile rate, measured for papillary muscle using the ‘zero-load clamp’ technique, is correlated with this isoenzymic change. Mechanisms associated with the onset of hypertrophy have been studied by continuous infusion of tyrosine C14 or lysine H3 as a protein synthesis precursor, and uridine H3 as an RNA precursor. While the rate of protein synthesis doubled after both aortic stenosis and aortic insufficiency, in the former case this doubling was seen after four days, while in the latter, it occurred only after 15 days. Myosin synthesis is normally twice as rapid as actin synthesis, and despite the increase in both, this proportion remained constant after four days of stenosis. The rise in protein synthesis preceded the increase in synthesis of ribosomal RNA 28 S fraction; thus the cardiac ribosomal stock is sufficiently large to allow an increase in protein synthesis in the absence of At novo ribosomal synthesis.

Protein synthesis as an early response to fertilization of the sea urchin egg: A model

We have developed a quantitative computer model which simulates the rise in protein synthesis resulting from the fertilization of the sea urchin egg. The model predicts the kinetics of incorporation of radioactively labeled amino acids into proteins for the experimental situation in which the amino acid pool is labeled prior to fertilization. The computer model is used to examine the impact of changes in the values of major parameters such as the time of initiation of protein synthesis, the rate at which mRNA is unmasked, the ribosome transit time, and the rate of depletion of the labeled amino acid pool on the kinetics of amino acid incorporation. When experimentally determined values for these parameters are used the model predicts kinetics which closely approximate the kinetics actually observed in newly fertilized eggs. We suggest that the rate at which mRNA is made available for translation and a change in the elongation rate following fertilization control the rise in protein synthesis, and that both of these processes are initiated within 0–2 min following the initial fertilization event.

Protein synthesis in resting and growth-stimulated human peripheral lymphocytes: Evidence for regulation by a non-messenger RNA

Stimulation of lymphocyte growth is accompanied by an early increase in the rate of protein synthesis. This increase is dependent upon the flow of inactive free ribosomes into polysomes, which is limited by a rate-controlling step at initiation [2]. Addition of actinomycin D (actD) to lymphocytes caused a gradual reduction in protein synthesis in resting cells, but rapidly inhibited both the elevation of protein synthesis and the activation of free ribosomes which normally follow exposure to mitogens. Since actD does not affect protein synthesis in enucleated lymphocytes [4], the effect in intact cells must be mediated by a nuclear event, which available data indicate is RNA synthesis. ActD prevented the accumulation of 80S initiation complexes which normally occurs in resting lymphocytes treated with pactamycin and cycloheximide, showing that its locus of action was at some point in initiation. The decline in rate of protein synthesis began without detectable lag when resting lymphocytes were treated with actD. However, after growth stimulation, a delay of ca 50 min occurred before the protein synthetic rate declined in response to actD. These observations agree with the hypothesis that the concentration of some moderately short-lived RNA is rate-limiting for protein synthesis in resting lymphocytes, and that an early event in growth stimulation is a rise in the amount of this component to levels which are no longer rate-limiting. This permits an increased flow of ribosomes into polysomes and a consequent rise in protein synthesis. Available evidence indicates that the regulatory RNA is neither mRNA nor rRNA, but may either be one of the small cytoplasmic RNAs whose function is unknown, or tRNA i met.

Effects of amino acids on cell differentiation of D. discoideum

The differentiation of D. discoideum amoebae is triggered by starvation, in particular, by starvation for amino acids. To examine the mechanism(s) by which amino acids inhibit cell differentiation we monitored the effects of specific groups of amino acids on protein synthesis and on amino acid catabolism, as judged by extracellular ammonia production. The experiments show that one group of 11 amino acids stimulates ammonia production and that this increase is dependent upon the concurrent rise in protein synthesis. A group composed of 7 other amino acids does not affect protein synthesis and elicits only a slight increase in ammonia production. In this case, the rise in ammonia does not require concommitant protein synthesis. These and other results suggest that specific groups of amino acids function to maintain the cells, metabolic activity, one by stimulating protein synthesis and the other by providing carbohydrates useful in energy production or other pathways, e.g., glycoprotein synthesis. The results are also consistent with the hypothesis that ammonia does not mediate the effects of amino acids on cell differentiation and probably does not play a regulatory role in the events leading to aggregation competence.