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G Link

Publications and source records attributed to G Link.

At least 55 records · Page 3Linked to original sources
PubMed1995

Sequence and expression characteristics of three G-box-binding factor cDNAs from Brassica napus [corrected].

G-box-binding factors (GBFs) are bZIP proteins that have been implicated in the transcriptional control of a number of plant genes including the family for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Using rbcS promoter regions as recognition site probes, we have cloned three Brassica GBFs designated as BnGBF1a, 1b and 2a. RNA gel blot analyses showed that all three BnGBF sequences give transcripts of the same size (1.3 kb) but in different amounts at a constant ratio in various tissues and developmental stages (1a > 2a > 1b). Transcript pools were largest in photosynthetically active organs such as leaves and cotyledons. Pool sizes correlated with those of total rbcS transcripts.

Amino Acid Sequence↗
PubMed1995

Identification and characterization of the Arabidopsis thaliana chloroplast DNA region containing the genes psbA, trnH and rps19'.

A 1887-nucleotide chloroplast-DNA region from Arabidopsis thaliana was analyzed. It contains the conserved genes psbA for the precursor of the D1 reaction-centre protein of photosystem II, trnH for tRNAHis, and rps19' for the 6.8-kDa protein of the small ribosomal subunit. Northern hybridization and RNase protection experiments suggest co-transcription of a minor RNA fraction over the full lengths of psbA and the preceding trnK-UUU gene, but not including downstream trnH sequences. In front of the mapped 5' end of the major 1.2-kb psbA transcript is a DNA region that shows the typical architecture of a psbA promoter, consisting of the prokaryotic-type '-35' and '-10' elements as well as the eukaryotic-type 'TATA' motif. The common 3' end of psbA transcripts seems to be located immediately after a stem-loop structure downstream from the coding region.

Arabidopsis↗
PubMed1995

Intrauterine vitamin B2 uptake of preterm and full-term infants.

Intrauterine uptake of vitamin B2 in preterm and full-term infants was examined. Factors of influence on vitamin supply were considered. Forty-four women and their infants were included in the study. Fetal vitamin uptake was calculated as arteriovenous concentration gradient in cord plasma times umbilical plasma flow. Concentration of vitamin B2 (free riboflavin and flavocoenzymes) was determined by high performance liquid chromatography of placental tissue and blood plasma (maternal vein, umbilical artery, umbilical vein). Flavocoenzymes were analyzed as flavin mononucleotide after acid hydrolysis of flavin adenine dinucleotide. Umbilical plasma flow was measured using pulsed Doppler sonography. Both free riboflavin and flavocoenzymes were transferred from the maternal plasma to the umbilical vein, but only free riboflavin was accumulated (approximately 1:4 for preterm and full-term infants, respectively). Flavocoenzyme concentration was higher in the umbilical vein than in the umbilical artery (p < 0.05). This indicated a median uptake of flavocoenzymes of 1.5 nmol/min.kg in preterm infants and 0.4 nmol/min.kg in full-term infants (preterm versus full-term, p < 0.01). Fetal vitamin supply depended on umbilical plasma flow and on maternal vitamin status (the latter was shown only in full-term infants). No dependence on placental vitamin concentration was observed (p > 0.05). Concentration of free riboflavin was higher in umbilical artery than in umbilical vein (p < 0.05). This indicated a release of free riboflavin from fetal tissues independent of gestational age (0.4 nmol/min.kg, preterm; 0.2 nmol/min.kg, full-term; p > 0.05).

Adult↗
PubMed1994

Ability of the orally effective iron chelators dimethyl- and diethyl-hydroxypyrid-4-one and of deferoxamine to restore sarcolemmal thiolic enzyme activity in iron-loaded heart cells.

In view of the profound functional and structural abnormalities shown in our previous studies in cultured, iron-loaded rat heart cells, we have examined the ability of the orally effective iron chelators dimethyl-3-hydroxypyrid-4-one (DMHP or L1) and diethyl-3-hydroxy-pyrid-4-one (DEHP or CP94) and of deferoxamine (DF) to reverse the damage caused by iron loading to heart cell organelles. At a concentration of 1.0 mmol/L, all three iron chelators were equally efficient in removing iron and restoring the activity of the thiolic sarcolemmal enzymes 5'-nucleotidase and Na,K,ATPase. However, at 0.1 mmol/L DMHP and DEHP were less effective than DF both in their iron-mobilizing effect and in promoting thiolic enzyme recovery. The superior efficiency of DF at low concentrations illustrates the advantage of the hexadentate chelating action of DF as compared with bidentate chelators such as DMHP and DEHP requiring a 3 to 1 molar ratio to iron for optimal effect. In contrast to its beneficial effect on sarcolemmal enzyme activity, iron chelation was unable to reverse the increase in beta-hexosaminidase activity caused by abnormal lysosomal fragility. Our study demonstrates for the first time that iron-induced peroxidative damage to the myocardial cell is associated with a marked loss of Na,K,ATPase activity, an enzyme with a major role in the maintenance of cellular resting potential. The timing of this damage and the restoration of Na,K,ATPase function by iron-chelating treatment suggest a cause-and-effect relationship between the observed injury to the sarcolemmal enzyme and the reversible electrophysiologic abnormalities observed in the same heart culture system in our previous studies.

5'-Nucleotidase↗
PubMed1994

Separation of two classes of plastid DNA-dependent RNA polymerases that are differentially expressed in mustard (Sinapis alba L.) seedlings.

Chloroplast and etioplast in vitro transcription systems from mustard have different functional properties, which is reflected in differences in phosphorylation status. Here we report another transcription control mechanism, which involves two plastid DNA-dependent RNA polymerases designated as peak A and peak B enzymes. Both are large multi-subunit complexes, but differ in their native molecular mass (> 700 kDa for peak A and ca. 420 kDa for peak B) and in their polypeptide composition. The A enzyme is composed of at least 13 polypeptides, while the B enzyme contains only four putative subunits. Peak B activity is inhibited by rifampicin, whereas that of peak A is resistant. RNA polymerase activity was compared for plastids from cotyledons of 4-day-old seedlings that were grown either under continuous light (chloroplasts) or in darkness (etioplasts), or were first dark-grown and then transferred to light for 16 h ('intermediate-type' plastids). While the total activity was approximately the same in all three cases, enzyme B was the predominant activity obtained from etioplasts and enzyme A that obtained from chloroplasts. Both had equal activity in preparations from the 'intermediate-type' plastid form. Both activation/inactivation and differential gene expression seem to play a role in the regulation of the plastid transcription machinery.

DNA-Directed RNA Polymerases↗
PubMed1993

Phosphorylation and dephosphorylation affect functional characteristics of chloroplast and etioplast transcription systems from mustard (Sinapis alba L.).

Chloroplast and etioplast RNA polymerase preparations each consist of a multi-subunit core and a set of three sigma-like transcription factors, SLF67, SLF52 and SLF29. Despite this structural similarity, the enzymes from either plastid type are functionally distinct, as is reflected by their different promoter usage and the tight core-SLF association in the etioplast but not the chloroplast holoenzyme. We tested whether these differences are related to phosphorylation. Treatment of the chloroplast enzyme with protein kinase converted it to an etioplast-type form and vice versa, treatment of the etioplast enzyme with phosphatase generated chloroplast-type properties. Although both the core enzyme and the SLF polypeptides were phosphorylation targets, only the SLFs seem to confer plastid-type-specific DNA binding characteristics. Methylation interference and DNase I footprint patterns in the psbA promoter region were found to correlate with the phosphorylation state of the chloroplast and etioplast enzymes.

Bacterial Proteins↗
PubMed1993

Sigma-like transcription factors from mustard (Sinapis alba L.) etioplast are similar in size to, but functionally distinct from, their chloroplast counterparts.

Three proteins resembling bacterial sigma factors were previously isolated from mustard chloroplasts (K. Tiller, A. Eisermann and G. Link, Eur J Biochem 198: 93-99, 1991). These sigma-like factors (SLFs) confer DNA-binding and transcription specificity to a system consisting of Escherichia coli core RNA polymerase and cloned DNA regions that carry a chloroplast promoter. Sigma-like activity was now isolated also from etioplasts and could be assigned to three polypeptides of M(r) 67,000 (SLF67), 52,000 (SLF52) and 29,000 (SLF29), i.e. the same sizes as for the chloroplast SLFs. The purification scheme for the factors from either plastid type included an initial heparin-Sepharose and a final gel filtration step. For the etioplast factors, however, an additional phosphocellulose step was required to release these polypeptides from the RNA polymerase. The etioplast SLFs have similar, but not identical, salt requirements for DNA binding as compared to their chloroplast counterparts. Under conditions of maximum binding activity there is overall preference of etioplast SLFs for the psbA promoter over the trnQ and rps16 promoters.

Base Sequence↗
PubMed1993

The 54 kDa RNA-binding protein from mustard chloroplasts mediates endonucleolytic transcript 3' end formation in vitro.

A 54 kDa protein from mustard chloroplasts was previously shown to interact specifically with a conserved U-rich sequence element in RNA derived from the 3' flanking regions of the plastid trnK and rps16 genes, which code for tRNA(Lys) and ribosomal protein CS19, respectively (Nickelsen and Link, 1991). This RNA-binding protein has now been purified by affinity chromatography on heparin Sepharose and poly(U) Sepharose. In vitro processing experiments and nuclease S1 analyses of the processing products revealed that the 54 kDa polypeptide is an endonuclease. The in vitro cleavage sites are consistent with the positions of corresponding transcript in vivo 3' ends downstream of trnK and rps16, suggesting that RNA 3' end formation takes place endonucleolytically also in vivo.

Base Sequence↗
PubMed1993

Measurement of 3-methylhistidine production in lambs by using compartmental-kinetic analysis.

The kinetics of 3-methylhistidine (3MH) metabolism in four crossbred lambs were studied. Each lamb was injected with an intravenous dose of 3-[2H3]methylhistidine (d3-3MH) and the stable isotope disappearance in plasma and appearance in both urine and muscle were measured. Immediately after the administration of tracer there was a phase of rapid disappearance of tracer from the plasma, which was followed by a more gradual decrease in d3-3MH from the plasma during the last 4 d of the experiment. A minimum of three exponentials was required to describe the plasma decay curve adequately. The kinetic model of 3MH in the whole animal was constructed by using the SAAM/CONSAM computer modelling program. Two different configurations of a three-compartment model are described: (1) a simple three-pool model, in which plasma kinetics were entered into pool 1 out of which they had one undefinable exit; (2) a plasma-urinary three-pool model with two exits, in which the urinary kinetics were entered as an exit out of pool 1 and required a second exit out of pool 3 to produce an adequate fit. In addition, muscle kinetics from biopsies of the longissimus dorsi were entered into either pool 2 or 3 using the plasma-urinary model. Steady-state mass and transport rate values were obtained for each model construct described, and a de novo production rate corresponding to a fractional breakdown rate of myofibrillar protein of approximately 5%/d was also calculated. The model predicted that only 15% of 3MH was excreted in urine as free 3MH, which is consistent with current knowledge of 3MH excretion in sheep. The simple three-pool plasma kinetic model, therefore, could be used to estimate, by a relatively simple injection-sampling technique, the extent of muscle protein turnover in lambs.

Animals↗
PubMed1993

The role of iron and iron chelators in anthracycline cardiotoxicity.

The redox cycling of anthracyclines promotes the formation of free radicals which are believed to play a central role in their cardiotoxicity. A number of observations indicate that the mechanism of the antineoplastic effect of anthracyclines is independent of their cardiotoxic effect and that it may be possible to prevent toxicity without interfering with therapeutic effect. Iron plays an important role in anthracycline toxicity by promoting the conversion of superoxide into highly toxic hydroxyl radicals through the Haber-Weiss reaction. Conversely, iron deprivation by its high-affinity binding to iron chelating compounds may inhibit anthracycline toxicity by interfering with free radical formation. ICRF-187, a bispiperazonedione which is hydrolyzed intracellularly into a bidentate chelator resembling EDTA, is able to decrease adriamycin-induced free hydroxyl radical formation and to prevent the development of clinical cardiac toxicity in patients receiving long-term anthracycline therapy. Our studies in rat heart cell cultures have shown that iron overload aggravates anthracycline toxicity and that this interaction can be prevented by prior iron chelating treatment. Since iron overload caused by multiple blood transfusions and bone marrow failure is a common condition in patients requiring anthracycline therapy, these observations may have significant clinical implications to the prevention of anthracycline cardiotoxicity.

Animals↗
PubMed1993

Anthracycline toxicity is potentiated by iron and inhibited by deferoxamine: studies in rat heart cells in culture.

The interrelation between iron, iron chelation, and anthracycline toxicity was investigated in a heart cell culture system. Two indicators of cellular damage have been used, lactate dehydrogenase (LDH) release and cell contractility. Both of these indicators have shown a marked increase in doxorubicin toxicity by prior iron loading. This was not a simple additive effect, because at the concentrations used, iron had only a minimal effect on LDH release and no effect at all on contractility, whereas doxorubicin had only a minor effect on contractility. Deferoxamine (DF) treatment of iron-loaded heart cells resulted in a marked decrease in anthracycline toxicity as judged both by LDH leakage and cell contractility. However, DF treatment of normal heart cells had no measurable protective effect against doxorubicin toxicity, whether DF was administered before or simultaneously with doxorubicin. Doxorubicin treatment did not alter cellular malondialdehyde (MDA) concentrations in either normal or iron-loaded cells. Conversely, the protective effect of DF in iron-loaded cells and its failure to prevent anthracycline toxicity in normal cells were both associated with a significant decrease in MDA measurements. Our data indicate that iron overload aggravates anthracycline toxicity and that this interaction may be prevented by effective iron chelating therapy. Because patients requiring anthracycline therapy often have increased tissue iron stores caused by multiple blood transfusions and bone marrow suppression, our observations may have important implications for the prevention of anthracycline toxicity.

Animals↗
PubMed1993

Iron loading of cultured cardiac myocytes modifies sarcolemmal structure and increases lysosomal fragility.

The mechanism of damage to myocardial subcellular organelles was studied in iron-loaded rat myocardial cells in culture in an attempt to identify the primary target of iron's toxic effects. Lysosomes and sarcolemmal membranes were purified by fractionation of the postnuclear supernatant on a 6.7% colloidal polyvinylpyrrolidone-coated silica gradient. After 24-hour incubation with ferric ammonium citrate at a concentration of 20 micrograms/ml (0.36 mmol/L) iron, a selective depletion of polyunsaturated fatty acids was found in whole-cell homogenates, as well as in the postnuclear supernatant and sediment. Iron loading resulted in a sharp increase in the total activity of the lysosomal enzyme beta-hexosaminidase in unfractionated whole-cell homogenates, increased free enzyme activity, and loss of latent activity indicating increased lysosomal fragility. Conversely, iron loading resulted in a marked decrease in the activity of the sarcolemmal enzyme 5'-nucleotidase and a significant loss of total protein sulfhydryl group content. These studies in cultured heart cells are in agreement with previous observations indicating increased lysosomal fragility in iron-loaded hepatic and splenic tissues, attributed to increased membrane lipid peroxidation. In addition, the marked decrease in sarcolemmal 5'-nucleotidase activity and in total protein sulfhydryl group content imply that iron-induced peroxidative damage to membrane proteins may be a more important mechanism in the pathogenesis of altered myocardial function in the iron-loaded heart than formerly was recognized.

5'-Nucleotidase↗
PubMed1992

Origin and fate of iron mobilized by the 3-hydroxypyridin-4-one oral chelators: studies in hypertransfused rats by selective radioiron probes of reticuloendothelial and hepatocellular iron stores.

The mechanism of in vivo iron chelation by 3-hydroxypyridin-4-ones (CP compounds) was studied in hypertransfused rats in which the major storage iron pools in hepatocytes and in the reticuloendothelial (RE) system have been labeled by selective radioiron probes. Both dimethyl-3-hydroxypyridin-4-one (CP 20 or L1) and diethyl-3-hydroxypyridine-4-one (CP 94) have an identical and very high (log beta 3 36) binding constant and selective affinity to iron(III), but the lipid solubility of CP 94 is considerably higher than that of CP 20. Both chelators induced an increase in the fecal excretion of hepatocellular iron with no effect on urinary excretion. In contrast, about one third to one half of the iron mobilized from RE cells was excreted in the urine. The chelating efficiency of CP 20 was comparable with that of deferoxamine (DF), whereas CP 94 was up to eight times more effective than DF. Unlike DF, which had no effect by the oral route, the oral and parenteral effectiveness of both CP compounds was identical. These findings indicate that: (1) lipid solubility is an important determinant of in vivo chelating efficiency; (2) urinary iron excretion induced by the CP compounds is derived from RE cells; (3) part of the iron mobilized from RE cells and all of the iron derived from hepatocytes is excreted through the bile; and (4) contrary to previous observations in cell cultures, there is no in vivo evidence for a diminishing chelating efficiency at the lowest doses used.

Animals↗