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Biomedical subjects

G Schuster

Publications and source records attributed to G Schuster.

At least 73 records · Page 4Linked to original sources
PubMed1989

Mutagenicity assay with Salmonella typhimurium revealing biotransformation of antiphytoviral substances by cell-free plant extract.

The mutagenic activity of four antiphytoviral substances was tested in reversion mutagenicity assays with a set of histidine auxotrophic strains of Salmonella typhimurium by means of the preincubation method. A possible metabolic activation of the substances by cell free fractions from maize seedlings (S-14-fraction) and for comparison from mouse liver (S-9 mix) was examined. None of the guanidine, phenyl urea and thiadiazole compounds exerted mutagenic activity in the bacterial strains in experiments without metabolic activation. Cyanoguanidine and N-phenyl-N-carboxyphenylurea became mutagenic for Salmonella strain TA98 after metabolic activation by the S-14 plant fraction. Both substances were not mutagenic in the presence of S-9 mix made from mouse liver. The promutagen cyclophosphamide proved highly mutagenic in experiments with S-14 mediated plant metabolic activation. This kind of bacterial mutagenicity assay is valuable in investigations of potential agrochemicals, as the examples have shown.

Animals↗
PubMed1988

Turnover of thylakoid photosystem II proteins during photoinhibition of Chlamydomonas reinhardtii.

The turnover of photosystem-II proteins during photoinhibition was analyzed in the green alga Chlamydomonas reinhardtii. Changes in the amount of photosystem II core complex polypeptides D1, D2, 44 kDa and 51 kDa, the antennae-CP-29 and light-harvesting-complex-II polypeptides and the water-oxidizing complex polypeptides of 30 kDa, 23 kDa and 16 kDa were monitored by a variety of techniques. Only the D1 and D2 polypeptides were found to turnover during photoinhibition when cells were exposed to ten fold photosynthesis-saturating light (2500 W/m2 for 90 min) at 25 degrees C. While 80% of photosystem-II activity was lost, a reduction of only 20% was observed in the total amount of D1 and D2 proteins. However, inhibition of chloroplast translation by chloramphenicol during photoinhibition resulted in the loss of about 60% of the D1 and 40% of the D2 proteins, as demonstrated by Western blotting and dot blotting of isolated thylakoids, quantitative analysis of immunogold-labeled whole-cell thin sections, and chase of radioactively prelabelled proteins during photoinhibition. We propose that the light-dependent turnover of the D1 protein is a protective mechanism against photoinhibition as far as the removal and replacement of D1 is compatible with the photoinactivation incurred by photosystem II. At light intensities at which the rate of D1 removal becomes limiting, loss of photosystem-II activity exceeds the turnover of D1 and the stability of the D2 protein is impaired as well.

Chlamydomonas↗
PubMed1988

Structure and biogenesis of Chlamydomonas reinhardtii photosystem I.

The photosystem I complex of the green alga Chlamydomonas reinhardtii was isolated and fractionated into its two subcomplex components: the core complex (CC I), which contained the reaction center (P-700) and had four polypeptide subunits, and the light-harvesting complex (LHC I) which contained four polypeptides of about 22, 25, 26 and 27 kDa. The 22-kDa apoprotein was isolated as a chlorophyll a and b binding protein. In the isolated photosystem I holocomplex, about ten copies of the 22-kDa LHC I apoprotein are present for each CC I unit. The 22-kDa polypeptide as well as the other three polypeptides of this complex and the subunit II of CC I are translated on 80S cytoplasmic ribosomes, and therefore are coded in the nucleus. During the greening process of the Chlamydomonas reinhardtii y-1 mutant the 22-kDa LHC I polypeptide, which cross-reacts with polyclonal antibodies raised against the Lemna gibba 20-kDa LHC I apoprotein, accumulates in thylakoids at a late stage of their development, and about 2-3 h after the LHC II and CC I subunit II polypeptides have accumulated. Accumulation of the 22-kDa protein during greening is inhibited by cycloheximide but not by chloramphenicol.

Chlamydomonas↗
PubMed1988

Role of the cytochrome b6.f complex in the redox-controlled activity of Acetabularia thylakoid protein kinase.

The regulation of the protein kinase activity responsible for the phosphorylation of the light-harvesting complex of photosystem II (LHCII) 27-kDa polypeptide involved in the State I-State II transitions in Acetabularia thylakoids was investigated. The LHCII kinase of isolated thylakoids retains its activity in absence of light-driven electron flow or reductants added in the dark. However, the kinase is reversibly inactivated by addition of oxidants in vitro or by far red (710 nm) light in vivo. Inhibitors of the quinol oxidase site of the cytochrome b6.f complex inactivate the LHCII kinase in the dark, and also in the light, or in presence of duroquinol when the plastoquinone pool is reduced. Inhibitors of the quinone reductase site of the b6.f complex have practically no effect in the dark and stimulate the kinase activity in the light. Based on these data and on our previous report, showing specific loss of LHCII kinase activity in a Lemna mutant lacking the cytochrome b6.f complex (Gal, A., Shahak, Y., Schuster, G., and Ohad, I. (1987) FEBS Lett. 221, 205-210), we propose that the activity of the LHCII kinase is regulated by the redox state of a cytochrome b6.f complex component(s) which responds to the balance of electron flow from photosystem II via the plastoquinone pool to photosystem I.

Acetabularia↗
PubMed1988

Evidence for protection by heat-shock proteins against photoinhibition during heat-shock.

The nuclear-coded 22 kd heat-shock protein (HSP-22) which is transported into the chloroplast and localized in the thylakoids was further characterized and found to be located in the grana lamellae (stacked thylakoids) as an extrinsic protein in the green alga Chlamydomonas reinhardtii. Inhibition of photosynthetic electron flow during heat-shock of Chlamydomonas cells was light-dependent, occurring at low-light intensities (<100 W/m) as compared with photoinhibition at 25 degrees C (>1000 W/m). The site of the damage was localized at the photosystem II (PS II) reaction center. The damage was drastically increased when heat-shock treatment was carried out in the presence of the 80S ribosomal translation inhibitor, cycloheximide (CHI). Pre-incubation of Chlamydomonas cells at 42 degrees C resulted in partial protection against photoinhibition during heat-shock, as compared with cells pre-incubated at 42 degrees C in the presence of CHI which, therefore, did not translate the heat-shock proteins. Analysis of the thylakoid polypeptides' pattern by SDS-PAGE revealed that during heat-shock in the light, thylakoid proteins became aggregated proportionally to the light intensity. Heat-shock in the presence of CHI enhanced the aggregation process which, at low light intensities, was specific to the PS II reaction center D1-protein. The results suggest that the chloroplasts HSPs prevent damage to the PS II reaction center during heat-shock in the light.

Journal Article↗
PubMed1988

Inhibitory effects of 9-(2,3-dihydroxypropyl)adenine and 3-(adenin-9-yl)-2-hydroxypropanoic acid 2-methylpropylester on potato virus X replication.

9-(2,3-Dihydroxypropyl)adenine (DHPA) and 3-(adenin-9-yl)-2-hydroxypropanoic acid 2-methylpropylester (AHPA-MP)) markedly inhibit the replication of potato virus X (PVX). The latter compound inhibits virus replication more effectively at the lower concentration range. In synchronized virus-infected leaf-disks, the time-response course of inhibition by both compounds is almost identical, and similar to that of cycloheximide.

Adenine↗
PubMed1987

Phosphorylation of spinach chlorophyll-protein complexes. CPII, but not CP29, CP27, or CP24, is phosphorylated in vitro.

Previous studies have indicated that the reversible phosphorylation of a population of antenna complexes that can donate energy to PS II ('mobile LHC II') plays a regulatory role in the state 1-state 2 transition in thylakoid membranes. The relationship of phosphorylated LHC II to the multiple PS II-associated chlorophyll a/b-proteins resolvable on green gels is currently unclear. We have used a high resolution gel system to analyze thylakoids phosphorylated in vitro. The only PS II-associated antenna complex to become phosphorylated is CPII, indicating that this complex represents the mobile LHC II. The other putative PS II antenna complexes, CP29, CP24, and the new complex designated CP27 which comigrates with CPII, are not phosphorylated and are probably components of the bound 'LHC II' antenna.

Chlorophyll↗
PubMed1987

Analogues of pyrimidine base precursors as antiphytoviral agents.

Dihydroorotic acid hydrazide (DHOH) did not inhibit the replication of potatovirus X (PVX) in leaf disks of Nicotiana tabacum 'Samsun'. In contrast, 5-fluoroorotic acid (5-FOA) completely inhibited the replication of PVX, as demonstrated by a serological virus assay as well as a local lesion bioassay using Gomphrena globosa as the test plant. The corresponding base analogue 5-fluorouracil (5-FU) had only a weak inhibitory effect. Time-course inhibition experiments in synchronized virus-infected leaf disks led us to conclude that 5-FOA, as well as 5-azadihydrouracil (5-ADHU), an uracil catabolite analogue, inhibit the same early event in virus infection. Neither 5-FOA nor 5-ADHU had a direct inactivating effect on free PVX virions.

Antiviral Agents↗
PubMed1986

Photosystem I reaction centers from maize bundle-sheath and mesophyll chloroplasts lack subunit III.

Photosystem I reaction centers were isolated from mesophyll and bundle-sheath chloroplasts of the C4 maize plant. Both preparations were found to be free of chlorophyll b and to have the same spectral properties and chlorophyll/P700 ratio as photosystem I reaction centers isolated from C3 plants. Photosystem I reaction centers from both mesophyll and bundle sheath were found to consist of six subunits with apparent molecular masses of about 70 kDa, 20 kDa, 17 kDa, 16 kDa, 10 kDa and 8 kDa, corresponding to photosystem I reaction center subunits I, II, IV, V, VI and VII of spinach, as tested by their immunological cross-reactivity with antibody raised against the respective spinach subunits. No cross-reactivity was found with antibodies raised against subunit III of spinach, either in whole thylakoids or purified reaction centers of both bundle-sheath and mesophyll chloroplasts. It is concluded that photosystem I reaction centers of bundle-sheath and mesophyll thylakoids of maize are identical and lack the polypeptide corresponding to subunit III present in all C3 plants so far tested.

Centrifugation, Density Gradient↗
PubMed1986

Transient inactivation of the thylakoid photosystem II light-harvesting protein kinase system and concomitant changes in intramembrane particle size during photoinhibition of Chlamydomonas reinhardtii.

Light-dependent reduction of the plastoquinone pool regulates the activity of the thylakoid-bound protein kinase which phosphorylates the light harvesting chlorophyll a,b-protein complex (LHC II) and regulates energy distribution between photosystems II (PS II) and I (Staehelin, L. A., and C. J. Arntzen, 1983, J. Cell Biol., 97:1327-1337). Since reduction of plastoquinone by PS II is abolished in photoinhibited thylakoids due to loss of the secondary electron acceptor QB protein (Kyle, D. J., I. Ohad, and C. J. Arntzen, 1984, Proc. Natl. Acad. Sci. USA, 81:4070-4074), it was of interest to examine the activity of the LHC II protein kinase system during photoinhibition and recovery of PS II activity. The kinase activity was assessed both in vivo and in vitro in Chlamydomonas cells exposed to high light intensity (photoinhibition) and recovery at low light intensity. The kinase activity was progressively reduced during photoinhibition and became undetectable after 90 min. The inactive LHC II-kinase system could not be reactivated in vitro either by light or by reduction of the plastoquinone pool following addition of reduced duroquinone (TMQH2). The LHC II polypeptides were dephosphorylated in vivo when cells, prelabeled with [32P]orthophosphate before exposure to high light intensity, were transferred to photoinhibiting light in the presence of [32P]orthophosphate. In vivo recovery of the LHC II-kinase activity, elicited by the addition of TMQH2 to the assay system, did not require restoration of QB-dependent electron flow or de novo protein synthesis, either in the cytoplasm or in the chloroplast. Mild sonication of thylakoids isolated from photoinhibited cells restored the ability of the LHC II protein kinase system to be activated in vitro by addition to TMQH2. Restoration of the light-activated LHC-II kinase required recovery of QB-dependent electron flow. At the structural level, photoinhibition did not affect the ratio of grana/stroma thylakoids. A reduction of approximately 20% of the 11-17-nm intramembrane particles and an equivalent increase in the number of 6-10.5-nm particles was observed on the E-fracture faces of stacked thylakoid membranes. Similar but smaller changes were observed also on the E-fracture faces of unstacked thylakoid membranes (more 10-14-nm and less 6-9-nm particles) and P-fracture faces of stacked thylakoid membranes (more 6-8- and less 9.5-13-nm particles). All these structural changes were reversed to normal values during recovery of PS II activity.(ABSTRACT TRUNCATED AT 400 WORDS)

Chlamydomonas↗
PubMed1986

Adaptation to CO(2) Level and Changes in the Phosphorylation of Thylakoid Proteins during the Cell Cycle of Chlamydomonas reinhardtii.

The photosynthetic performance of synchronously grown Chlamydomonas reinhardtii alternated rhythmically during the cell cycle. The activity of the "CO(2) concentrating mechanism" including the ability to accumulate CO(2) internally and the activity of carbonic anhydrase peaked after 6 to 9 hours of light and reached minimum after 6 to 9 hours of dark. Consequently, the apparent photosynthetic affinity to extracellular CO(2) alternated rhythmically. At the end of the dark period the cells behaved as if they were adapted to high CO(2) even though they were continuously aerated with air. Results from experiments in which the light or dark periods were extended bear on the interaction between the internal (cell cycle or biological clock) and the external (light) signal. The observed rhythmical alterations in photosynthetic V(max) may result from changes in PSII activity. The latter may be partly explained by the capacity for phosphorylation of thylakoid proteins, which reached maximum after 9 hours of light and decreased toward the dark period.

Journal Article↗
PubMed1985

Differentiation and development of bundle sheath and mesophyll thylakoids in maize. Thylakoid polypeptide composition, phosphorylation, and organization of photosystem II.

Photosynthetic electron flow, polypeptide pattern, presence of chlorophyll-protein complexes, and phosphorylation of thylakoid polypeptides have been investigated in differentiated mesophyll (M) and bundle sheath (B) thylakoids of the C4 plant Zea mays. The polypeptide pattern of M thylakoids and their photosynthetic electron flow are comparable to those of other green plants. B thylakoids exhibit only photosystem I (PSI) activity, contain only traces of the PSII light harvesting (LHCII) polypeptide, do not bind [3H] diuron, and lack polypeptides of the water-oxidation complex of PSII and the herbicide binding 32-kDa polypeptide, as detected by specific antibodies. However, B thylakoids possess a partially active PSII reaction center, as demonstrated by light-dependent reduction of silicomolybdate with 1,5-diphenylcarbazide (DPC) as an electron donor, and the presence of the PSII reaction center polypeptides of 44-47 kDa. Only one chlorophyll a-protein complex, corresponding to the PSI reaction center-core antenna, was detectable in B thylakoids, as opposed to chlorophyll a and chlorophyll a,b-protein complexes present in M thylakoids. The light-dependent, membrane-bound kinase activity present in M thylakoids could not be detected in B thylakoids which, nevertheless, contain a protein kinase able to phosphorylate casein. A total of 19 differences between the electrophoretic pattern of B and M thylakoid polypeptides were observed. The mRNA coding for the LHCII polypeptide is primarily, if not exclusively, localized in M cells. The development of PSII complex precedes that of PSI during the differentiation of B and M chloroplasts in expanding leaves of light-grown plants and during the greening of dark-grown etiolated seedlings. The differentiation of the maize leaf into cells programmed to form B or M chloroplasts does not require light. In light-grown plants, the differentiation of B and M thylakoids occurred progressively from the base of the leaf and was completed at 4-5 cm from the leaf base.

Cell Differentiation↗
PubMed1985

[Cancer of the operated stomach].

On the basis of the own documents of patients and the documents of the post-mortem examination the increased risk of carcinoma of the resected stomach is confirmed. Extensive experiments on the rat seem to ascribe the greatest role in the etiology of this special form of gastric cancer to the lesion of the mucous by the bile reflux and the settlement of the operated stomach with nitrate-reducing germs.

Adenocarcinoma↗
PubMed1985

Synthesis, transport and localization of a nuclear coded 22-kd heat-shock protein in the chloroplast membranes of peas and Chlamydomonas reinhardi.

The synthesis, transport and localization of a nuclear coded 22-kd heat-shock protein (HSP) in the chloroplast membranes was studied in pea plants and Chlamydomonas reinhardi. HSPs were detected in both systems by in vivo labeling and in vitro translation of poly(A)RNA, using the wheat-germ and reticulocyte lysate systems. Heat-shock treatment of pea plants for 2 h at 42-45 degrees C induces the expression of 10 nuclear coded proteins, among which several (18 kd, 19 kd, 22 kd) are predominant. A 22-kd protein is synthesized as a 26-kd precursor protein and is localized in a chloroplast membrane fraction in vivo. Following post-translational transport into intact chloroplasts in vitro of the 26-kd precursor, the protein is processed but the resulting 22-kd mature protein is localized in the chloroplast stroma. If, however, the in vitro transport is carried out with chloroplasts from heat-shocked plants, the 22-kd protein is preferentially transported to the chloroplast membrane fraction. In C. reinhardi the synthesis of poly(A)RNAs coding for several HSPs is progressively and sequentially induced when raising the temperature for 1.5 h from 36 degrees C to 42 degrees C, while that of several preexisting RNAs is reduced. Various pre-existing poly(A)RNAs endure in the cells at 42 degrees C up to 5 h but are no longer translated in vivo, whereas some poly(A)RNAs persist and are translated. As in pea, a poly(A)RNA coded 22-kd HSP is localized in the chloroplast membranes in vivo, although it is translated as a 22-kd protein in vitro. The in vitro translated protein is not transported in isolated pea chloroplast which, however, processes and transports other nuclear coded chloroplast proteins of Chlamydomonas. The poly(A)RNA coding for the 22-kd HSP appears after 1 h at 36 degrees C. Its synthesis increases with the temperature of incubation up to 42 degrees C, although it decreases after 2 h of heat treatment and the already synthesized RNA is rapidly degraded. The degradation is faster upon return of the cells to 26 degrees C. None of the heat-induced proteins is identical to the light-inducible proteins of the chloroplast membranes.

Journal Article↗
PubMed1984

Mode of action of the antiphytoviral compound 2,4-dioxohexahydro-1,3,5-triazine (5-azadihydrouracil).

The time course of inhibition of potato virus X ( PVX ) synthesis by the newly developed antiphytoviral compound 2,4- dioxohexahydro -1,3,5-triazine (DHT) was determined in mechanically inoculated leaves of Nicotiana tabacum L. cv. ' Samsun '. At the permissive temperature (22 +/- 3 degrees) DHT inhibited the synthesis of PVX almost 100% within 8 h postinoculation, after which the inhibition declined. In leaves maintained at a temperature of 5 +/- 2 degrees for 5 days, DHT inhibited PVX synthesis about 65% immediately after the shift to permissive temperature. This time course of inhibition was almost identical to that of 2-thiouracil. Uracil completely overcame the inhibition of PVX replication caused by DHT when added within 2 h after incubation of leaf disks with DHT. These results indicate that DHT acts as an analogue of the pyrimidine base uracil.

Antiviral Agents↗
PubMed1984

[Degradation and utilization of 2,4-dioxohexahydro-1,3,5-triazine (DHT) by soil microorganisms].

The biodegradation and utilization of the antiphytoviral substance 2,4-dioxohexahydro-1,3,5-triazine (DHT) by soil microorganisms was investigated. Mixed cultures of microorganisms deriving from different soils diminish in nutrient broth the content of DHT with increasing duration of culture. Microorganisms from an Egyptian garden soil fully degrade 10(-3) mol/1 DHT in a culture without additional aeration within 28 days. Also in deficient media the mixed microorganisms reduce the amount of DHT, reaching in nitrogen free nutrient solution even a degradation rate up to 12 mg DHT per liter and day. Pure cultures of Rhizobium leguminosarum, Proteus vulgaris, Saccharomyces cerevisiae and especially Agrobacterium radiobacter diminish the content of DHT in nitrogen free media, too. No such effect was detectable in cultures of four other species of soil bacteria. The DHT degradation by the microorganisms is connected with significant cell multiplication, e.g. A. radiobacter in shaking cultures with DHT as sole source of nitrogen shows a typical growth cycle with a lag-phase of 24 hours. The short persistence time of DHT in soils is concluded to be mainly due to biodegradation by microorganisms.

Antiviral Agents↗