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  • 1
    ISSN: 1432-2048
    Keywords: Nitrate-reductase mRNA ; Photooxidation of plastids ; Plastidic factor ; Sinapis (nitrate-reductase mRNA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It was found previously that in the mustard (Sinapis alba L.) seedling (Schuster et al. 1989, Planta 177, 74–83) the action of nitrate and phytochrome on the appearance of cytosolic nitrate reductase (NR) is abolished if the plastids are damaged by photooxidation. In the present study this finding has been corroborated by the following results: (i) the appearance and disappearance of NR activity are strictly correlated with the appearance and disappearance of immunoresponsive NR protein; (ii) the appearance of NR correlates with the appearance of translatable NR mRNA; (iii) photodestruction of the plastids strongly reduces the level of NR mRNA. It is concluded that the dependence of the NR level on the state of the plastids can be detected at the level of its mRNA and is not attributable to an inactivation of the enzyme.
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  • 2
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Nitrate (as inductor) ; Nitrite reductase ; Phytochrome (as inductor) ; Sinapis (nitrate assimilation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The extent to which the appearances of nitrite reductase (NIR; EC 1.7.7.1) and glutamine synthetase (GS; EC 6.3.1.2) are coordinated was studied in mustard (Sinapis alba L.) seedlings. It was established by immunotitration that the increased activities of NIR and GS in the presence of light and nitrate can be attributed to the de-novo synthesis of enzyme protein. The bulk of the NIR and GS was found in the developing cotyledons. In the absence of nitrate in the growth medium there was no coordinate appearance of NIR and GS. While light strongly stimulated the appearance of GS, the level of NIR was hardly affected and remained low. On the other hand, in the presence of nitrate in the medium the appearances of NIR and GS were strictly coordinated, the GS level being considerably above that of NIR. It is argued that phytochrome-controlled synthesis of GS in the absence of nitrate is part of the mechanism to reassimilate ammonium liberated during proteolysis of storage protein and metabolism of the resulting amino acids, whereas the strictly coordinated synthesis in the presence of light and nitrate indicates the dominance of nitrate assimilation under these circumstances. The fact that the level of GS was always considerably above that of NIR appears to be a safety measure to prevent ammonium accumulation.
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  • 3
    ISSN: 1432-2048
    Keywords: Gene expression (control points) ; Nitrate ; Nitrite reductase ; Phytochrome ; Sinapis (phytochrome)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrate reductase (NR, EC 1.6.6.1) and nitrite reductase (NIR, EC 1.7.7.1) are the key enzymes of nitrate reduction. It is well established that the appearance of these enzymes is “induced” by nitrate, and it is generally believed that NR is cytosolic while NIR is plastidic. In mustard (Sinapis alba L.) cotyledons we observed two isoforms of NIR (NIR1 and NIR2) using a chromato-focusing technique. Only one of them (NIR2) disappeared when the plastids were damaged by photooxidation in the presence of Norflurazon. It is concluded that NIR2 is plastidic while NIR1 is extraplastidic and not affected by photooxidation of the plastids. Both isoforms appear to have the same molecular weight (60 kilodaltons, kDa). Two distinct translation products which could be immunoprecipitated with NIR antiserum produced against total NIR from mustard were observed which differed slightly in molecular weight (60 versus 63 kDa). The 63-kDa polypeptide was considered to be the precursor of NIR2. While synthesis of NIR protein depended largely on nitrate, the levels of in-vitro-translatable NIR mRNAs were found to be either independent of nitrate and light (NIR1) or controlled by phytochrome only (NIR2). It appears that phytochrome strongly stimulates the level of mRNA while significant enzyme synthesis (NIR2) takes place only in the presence of relatively large amounts of nitrate. Since an increased enzyme level was strictly correlated with an increase of immunoresponsive NIR protein it is improbable that activation of a precursor plays a role. Rather, it is concluded that, in situ, nitrate controls translation.
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  • 4
    ISSN: 1432-2048
    Keywords: Gene expression ; Nitrate ; Nitrite reductase (control) ; Phytochrome ; Spinacia (nitrite reductasegene expression)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It is well established that nitrite reductase (NIR; EC 1.7.7.1) a key enzyme of nitrate reduction — is “induced” by nitrate and light. In the present study with the spinach (Spinacia oleracea L.) seedling the dependency of NIR appearance on nitrate, light and a ‘plastidic factor’ was investigated to establish the nature of the coaction between these controlling factors. A cDNA clone coding for spinach NIR was available as a probe. The major results we have obtained are the following: (i) The light effect on the appearance of NIR activity occurs through phytochrome. No specific bluelight effect is involved, (ii) Immunotitration data indicate that light affects the appearance of NIR by inducing the de-novo synthesis of the NIR protein, (iii) A multiplicative relationship exists between the action of nitrate and light on NIR appearance. This indicates that the actions of light and nitrate are indeed independent of each other but that both factors operate on the same causal sequence, (iv) Anion-exchange chromatography revealed only a single form of NIR in spinach. Experiments involving plastid photooxidation indicate that this NIR is exclusively plastidic. (v) Northern blot analysis of NIR mRNA showed a strong increase of the steady-state level in the presence of nitrate whereas light had no effect; NIR mRNA was almost undetectable when the plastids were damaged by photooxidation. It is concluded that NIR gene expression in spinach requires positive control by a ‘plastidic factor’. Moreover, nitrate exerts a coarse control at the mRNA level whereas fine tuning of NIR protein synthesis is post-transcriptional and is exerted by light, operating via phytochrome.
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  • 5
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Nitrate (as inductor) ; Nitrite reductase ; Phytochrome (as inductor) ; Sinapis (nitrate assimilation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The extent to which the appearances of nitrite reductase (NIR; EC 1.7.7.1) and glutamine synthetase (GS; EC 6.3.1.2) are coordinated was studied in mustard (Sinapis alba L.) seedlings. It was established by immunotitration that the increased activities of NIR and GS in the presence of light and nitrate can be attributed to the de-novo synthesis of enzyme protein. The bulk of the NIR and GS was found in the developing cotyledons. In the absence of nitrate in the growth medium there was no coordinate appearance of NIR and GS. While light strongly stimulated the appearance of GS, the level of NIR was hardly affected and remained low. On the other hand, in the presence of nitrate in the medium the appearances of NIR and GS were strictly coordinated, the GS level being considerably above that of NIR. It is argued that phytochrome-controlled synthesis of GS in the absence of nitrate is part of the mechanism to reassimilate ammonium liberated during proteolysis of storage protein and metabolism of the resulting amino acids, whereas the strictly coordinated synthesis in the presence of light and nitrate indicates the dominance of nitrate assimilation under these circumstances. The fact that the level of GS was always considerably above that of NIR appears to be a safety measure to prevent ammonium accumulation.
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  • 6
    ISSN: 1432-2048
    Keywords: Gene expression ; Nitrate ; Nitrate reductase ; Phytochrome ; Spirodela ; Turion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Control by light and nitrate of the appearance of nitrate reductase (NR; EC 1.6.6.1) in the turions of Spirodela polyrhiza (L.) Schieiden was investigated during the pre-germination phase, i.e. up to 120 h after the transfer from after-ripening to germination conditions. Turions are particularily suited for this type of coaction study since control of nitrate uptake by light and long-distance transport of nitrate do not play a role. Control of NR gene expression was mainly studied between 72 and 120 h after transfer to germination conditions (phase II of the pre-germination process). It was found that the effect of light on NR appearance is exerted via phytochrome. The light effect on enzyme synthesis was only observed in the presence of nitrate. On the other hand, in darkness, the NR level decreased to almost zero even in the presence of an optimum supply of nitrate. It is concluded that in phase II a coaction of light (phytochrome) and nitrate is required to bring about NR synthesis. However, when nitrate was applied to turions — following a dark incubation without nitrate — the turions responded to nitrate even in darkness with strong NR synthesis. This response was augmented by light. The highest transcript levels were observed in the presence of both factors, light and nitrate. As a single factor, light was more effective in stimulating the transcript level than nitrate. However, no correlation between the transcript level and the rate of enzyme synthesis was observed. This is consistent with the previous conclusion (Schuster et al. 1988, Planta 174, 426–432) that in higher plants control at the transcript level is only coarse and does not determine quantitatively the output at the level of enzyme protein.
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  • 7
    ISSN: 0021-8383
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Products of the Decarboxylation of Photochemically prepared Indenecarboxylic Acid Derivatives.4-(2-Diazo-1-oxo-1, 2-dihydro-naphthalen-5-sulfonyl-oxy)-benzophenone (NQD-BP) was photolyzed to form 4-(3-carboxy-indene-7-sulfonyl-oxy)-benzophenone (ICA-BP). This was decarboxylated base-catalysed in ethanol leading to 4-(indene-4-sulfonyl-oxy)-benzophenone (4) and 4-(indene-7-sulfonyl-oxy)-benzophenone (5). The indene isomers were separated by means of preparative HPLC and their structure was determined using n.m.r., mass, i.r. and u.v. spectroscopy.
    Additional Material: 4 Ill.
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  • 8
    ISSN: 0941-1216
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Different electronic structures for the three 1,2-naphthoquinone-2-diazide (NQD) chromophores in 2,3,4-tri (1,2-naphthoquinone-2-diazide-5-sulfonyl-oxy)-benzophenone were detected using 1H NMR spectroscopy. The NQD substituent in the o-position of the benzophenone is influenced by intramolecular through space interactions with the benzophenone part of the molecule. The change in the electronic structure is the reason for the obtained different photoreactivity of the o-NQD chromophore compared with m-NQD and p-NQD. The relative quantum yields of photolysis are 0.3 (o-NQD) and 0.9/1.0 (m-NQD/p-NQD).
    Additional Material: 2 Ill.
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  • 9
    ISSN: 0009-286X
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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