Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.27.5 (RNase)
17,967 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We cloned a cDNA for a gibberellin-induced ribonuclease (RNase) expressed in barley (Hordeum vulgare) aleurone and the gene for a second barley RNase expressed in leaf tissue. The protein encoded by the cDNA is unique among RNases described to date in that it contains a novel 23-amino acid insert between the C2 and C3 conserved sequences. Expression of the recombinant protein in tobacco (Nicotiana tabacum) suspension-cultured protoplasts gave an active RNase of the expected size, confirming the enzymatic activity of the protein. Analyses of hormone regulation of expression of mRNA for the aleurone RNase revealed that, like the pattern for alpha-amylase, mRNA levels increased in the presence of gibberellic acid, and its antagonist abscisic acid prevented this effect. Quantitative studies at early times demonstrated that cycloheximide treatment of aleurone layers increased mRNA levels 4-fold, whereas a combination of gibberellin plus cycloheximide treatment was required to increase alpha-amylase mRNA levels to the same extent. These results are consistent with loss of repression as an initial effect of gibberellic acid on transcription of those genes, although the regulatory pathways for the two genes may differ.
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PMID:Cloning and characterization of a gibberellin-induced RNase expressed in barley aleurone cells. 1019 5

Pale and homogeneous-looking nuclei of degenerative acinar cells selectively seen in an early regression stage of the human lactating breast were periodic acid-Schiff (PAS)-reactive. In our preceding paper, this peculiar morphologic feature was designated as 'magentosis'. The present paper was aimed at histochemically clarifying the nature of the 'magentotic' nuclei. The diffuse PAS reactivity was not influenced by pretreatments with alpha-amylase, DNase, RNase, proteinase K, nor by hydrochloric acid or heating. The nuclei were negative for acid mucosubstances and secretory glycoproteins, and were unreactive with a variety of lectins. In contrast, the presence of single-stranded DNA stretches or breaks was proven. The 'magentotic' nuclei in non-heated paraffin sections were hybridized with a heat-denatured DNA probe for human DNA consensus sequences and were focally immunoreactive with an antibody to single-stranded DNA. The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method turned to be positive after digestion by mung bean nuclease, a single-stranded DNA-specific enzyme. The 'magentotic' nuclei were further clearly labeled by the in situ nick translation method. The nucleoli were devoid of reactivity for both the PAS and single-stranded DNA signals. We propose that 'magentosis' represents a unique mode of cell death, distinct from apoptosis and necrosis or oncosis. The PAS reactivity in the 'magentotic' nuclei may be correlated with the occurrence of single-stranded stretches or breaks in the DNA chain.
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PMID:'Magentosis' in human lactating breast: a mode of cell death accumulating single-stranded DNA stretches or breaks. 1084 59

Among the physical factors which might influence micro-organisms one of the most potentially interesting is high vacuum. The effect of high vacuum is less studied as compared with other physical factors. It is impossible to achieve, under laboratory conditions, a vacuum of the order 10(-16) mm Hg which is probably characteristic of space. Earlier, the effect of high vacuum was studied on different bacteria, yeasts, molds and algae. It appeared that spores and fungal conidia were not killed by high vacuum. Later, the effect of high vacuum on physiological processes in micro-organisms was studied. The ability to oxidize glucose or ethanol was studied with Sarcina flava and Bacillus simplex cells after they were subjected for 72 hr to vacuum (10(-8) to 10(-9) mm Hg). The oxidation rate was followed polarographically. The oxidative ability of S. flava cells diminished [correction of dimished] after their subjection to vacuum, while B. simplex spores were unchanged in that respect. The following crystalline enzymes were subjected for 72 hr to the same vacuum: alpha-amylase, catalase, ribonuclease, trypsine and urease. Then the activity of the above enzymes was tested on corresponding substrates. Not a single enzyme was totally inactivated. About 50% of activity was lost with alpha-amylase; 25--35% of activity with catalase, ribonuclease and urease. Trypsine retained its total activity. Thus, high vacuum cannot be listed among factors rapidly inactivating enzymes of micro-organisms.
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PMID:The effect of high vacuum on oxidative reactions in bacteria and the activity of certain enzymes. 1266 21

1. Extracellular ribonuclease is produced linearly for at least 3hr. by washed post-logarithmic-phase cells of Bacillus subtilis suspended in a medium containing maltose (1%) and casein hydrolysate (0.5%). 2. Low concentrations of actinomycin D (less than 2mug./ml.) stimulate ribonuclease formation, the maximum effect being observed with a concentration of 1mug./ml. Concentrations greater than 2mug./ml. are inhibitory. There is no parallel stimulation of alpha-amylase formed under the same conditions, and [(14)C]uracil incorporation into a perchloric acid-insoluble form is inhibited. 3. The actinomycin D-induced stimulation is not due to the presence of an activator, nor is the inhibition due to the release of an inhibitor by the cells. The effect is on the amount of ribonuclease produced in the medium. 4. Extracellular ribonuclease formation is partially inhibited by anaerobiosis, 2,4-dinitrophenol, sodium azide and by chloramphenicol and puromycin. 5. High concentrations of antibiotic do not completely inhibit ribonuclease formation, but a basal amount of enzyme representing 20min. synthesis in an uninhibited system is always produced. This ;antibiotic-insensitive' enzyme could possibly represent preformed enzyme ;in the pipe-line' en route to secretion. 6. The stimulated appearance of ribonuclease in the presence of 1mug. of actinomycin D/ml. is shown to be dependent on enzyme synthesis. The mechanism of this effect is discussed.
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PMID:EXTRACELLULAR RIBONUCLEASE FORMATION IN BACILLUS SUBTILIS AND ITS STIMULATION BY ACTINOMYCIN D. 1434 5

Towards a goal of detecting scaled-up DNA adducts as altered deoxynucleotides by mass spectrometry, we have set up a practical and general method for isolating DNA-derived deoxyribonucleoside-5'-monophosphates devoid of ribonucleotides starting with a 1 g sample of mammalian tissue. The method is practical because costs have been minimized, and it is general because it can be applied to a more difficult sample such as mouse skin or non-fresh calf liver. The procedure, consisting of a series of steps that were largely gleaned and tuned from prior literature, proceeds as follows: (1) homogenize the tissue in sodium dodecyl sulfate; (2) digest with ribonuclease A, ribonuclease TI, alpha-amylase and proteinase K; (3) partition between water and phenol; (4) precipitate the DNA with ethanol followed by redissolving and dialysis; and (5) digest with nuclease P1 and phosphodiesterase I followed by ultrafiltration and boric acid gel chromatography. The yellow to brown color of DNA from difficult tissues only persisted up to the ultrafiltration step. Apparently this DNA was contaminated with iron-containing proteins. Residual ribonucleotides were not observable (<0.1%) by HPLC in the final sample. Without boric acid gel chromatography, residual contamination by ribonucleotides was about 1% even when the DNA was purified before digestion by phenol partitioning followed by use of a Genomic Tip kit from Qiagen.
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PMID:Phenolic extraction of DNA from mammalian tissues and conversion to deoxyribonucleoside-5'-monophosphates devoid of ribonucleotides. 1554 80

Gibberellic acid enhances the synthesis of alpha-amylase in isolated aleurone layers of barley-seeds (Hordeum vulgare var. Himalaya). In the presence of 20 mm calcium chloride the amount of enzyme obtained from isolated aleurone layers is quantitatively comparable to that of the half-seeds used in earlier studies. After a lag period of 6 to 8 hours enzyme is produced at a linear rate. Gibberellic acid does not merely trigger alpha-amylase synthesis, but it is continuously required during the period of enzyme formation. Enzyme synthesis is inhibited by inhibitors of protein and RNA synthesis. Small amounts of actinomycin D differentially inhibit enzyme release and enzyme synthesis suggesting 2 distinct processes. Gibberellic acid similarly enhances the formation of ribonuclease which increases linearly over a 48 hour period. During the first 24 hours the enzyme is retained by the aleurone cells and this is followed by a rapid release of ribonuclease during the next 24 hour period. The capacity to release the enzyme is generated between 20 and 28 hours after the addition of the hormone. Ribonuclease formation is inhibited by inhibitors of protein and RNA synthesis. These inhibitors also prevent the formation of the release mechanism if added at the appropriate moment.
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PMID:Gibberellic Acid-enhanced synthesis and release of alpha-amylase and ribonuclease by isolated barley and aleurone layers. 1665 17

Gibberellic acid (GA) enhances the synthesis of alpha-amylase and ribonuclease in isolated aleurone layers and this process is inhibited by abscisin. Removal of gibberellic acid in mid-course of alpha-amylase production results in a slowing down of alpha-amylase synthesis, suggesting a continued requirement of GA for enzyme synthesis. This is paralleled by a continuous requirement for RNA synthesis. Addition of 6-methylpurine or 8-azaguanine in mid-course results in an inhibition of alpha-amylase synthesis within 3 to 4 hours. However, actinomycin D added in mid-course is almost without effect. This is not due to its failure to enter the cells, because it does inhibit (14)C-uridine incorporation at this stage. Addition of abscisin to aleurone layers which are synthesizing alpha-amylase results in an inhibition of this synthesis within 2 to 3 hours. Cycloheximide on the other hand inhibits enzyme synthesis immediately upon its addition. These data are consistent with the hypothesis that the expression of the GA effect requires the synthesis of enzyme-specific RNA molecules. The similarity in the kinetics of inhibition between abscisin on the one hand and 8-azaguanine or 6-methylpurine on the other suggests that abscisin may exert its action by inhibiting the synthesis of these enzyme-specific RNA molecules or by preventing their incorporation into an active enzyme-synthesising unit.
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PMID:Hormonal control of enzyme synthesis: on the mode of action of gibberellic Acid and abscisin in aleurone layers of barley. 1665 90

Protector-II (Pr-II) of the Japanese morning glory (Pharbitis nil Choisy) was inactivated by exposure to polyphenol oxidase. An unidentified protector in the same molecular weight range obtained from sunflower was also inactivated by this enzyme. Earlier speculations that protectors might be lipoprotein in nature were negated by the fact that neither lipase nor protease inactivated the protectors. The protectors were also not inactivated by incubating with alpha-amylase, DNase, or RNase. Catechol mimics Pr and is inactivated by polyphenol oxidase. The oxidation of catechol to o-quinone is accompanied by a loss of chromophores that absorb ultraviolet light and the appearance of a reddish brown color. Similarly, when the relatively low molecular weight auxin protectors (Pr-II class) were incubated with polyphenol oxidase, their oxidation was also frequently associated with the formation of brown color, and oxidation with H(2)O(2) caused a loss of ultraviolet-absorbing chromophores. The data indicate that auxin protectors contain o-dihydroxyphenolic groups at their active site.That o-dihydroxyphenols inhibit indoleacetic acid oxidation has been demonstrated by numerous workers. It is suggested that the high molecular weight auxin protectors and the phenolic compounds described by other authors comprise part of a metabolic system concerned with the regulation of peroxidase-catalyzed redox reactions.
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PMID:Studies on Auxin Protectors: IX. Inactivation of Certain Protectors by Polyphenol Oxidase. 1665 85

Seeds of Dixie crimson clover (Trifolium incarnatum cv. Dixie) completed germination in 36 hours at 20 C. At 10 C germination was delayed by 24 hours. At 30 C only 20% germinated and the rest remained viable for a long time but not germinable. Different patterns of adenylate energy state and zymograms of acid phosphatase and esterase were observed from seeds grown under the three temperatures for 24 hours. Varied specific activities of protease, alpha-amylase, ATPase, RNase, acid phosphatase, glutamine synthetase, and fumarase were also found. Protein-synthesizing ability was proportional to temperature. These data indicate that temperature regulates seed germination at multiple sites.
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PMID:Temperature regulation of germination in crimson clover seeds. 1665 91

A rice cell suspension culture system with the Ramy3D promoter, which is induced by sucrose starvation, has been previously utilized to produce large quantities of recombinant proteins. Although this expression system was reported previously to generate a good yield of recombinant hGM-CSF in transgenic rice cell suspension culture, rice alpha-amylase was a dominant protein, with 43% of total secreted proteins and an obstacle to the production and purification of secreted recombinant proteins in a rice cell suspension culture. In this study, an intron-containing self-complementary hairpin RNA (ihpRNA)-mediated post transcriptional gene silencing (PTGS) strategy for the rice alpha-amylase gene was applied in order to overcome this problem in rice cell suspension culture systems. The reduction of the mRNA level of the rice alpha-amylase gene was verified via Northern blot analysis and siRNA, an initiator of RNA interference, was detected via an RNase protection assay. The amount of rice alpha-amylase in the culture medium was reduced to 8.2% as compared to that of the wild-type. A transgenic rice cell suspension culture expressing both the hGM-CSF and ihpRNA of the rice alpha-amylase gene demonstrated that the quantity of rice alpha-amylase was reduced to 22% and that the accumulation of hGM-CSF increased by 1.9-fold as compared to that in the transgenic cell line expressing hGM-CSF only. These results indicated that RNAi technology should be of great utility for suppressing undesirable genes, and should improve accumulation and facilitate the purification of secreted recombinant proteins in rice cell suspension cultures.
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PMID:Amylase gene silencing by RNA interference improves recombinant hGM-CSF production in rice suspension culture. 1863 17


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