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W D Sutton

Publications and source records attributed to W D Sutton.

15 recordsLinked to original sources
PubMed1979

Polypeptide synthesis by Rhizobium bacteroids and bacteria.

When Rhizobium bacteroids (strain NZP 2257) from lupin nodules were isolated and incubated aerobically at high osmolarity, they incorporated [35S]-methionine into a characteristic set of polypeptides; many of these polypeptides coelectrophoresed on SDS-polyacrylamide gels with the bacteroid polypeptide bands stained by Coomassie blue. The labelled polypeptides were stable for several hours in pulse-chase experiments. Changes in the concentration of H+, K+ and Mg2+ in the incubation mixture affected overall incorporation of label, but not the relative incorporation into different polypeptides. A similar set of bacteroid polypeptides was labelled in situ when detached nodules were fed [35S]methionine. Distinctive labelling patterns were observed with bacteroid suspensions from mature and immature nodules, with a transitional pattern at the time when nitrogenase activity appeared. Two of the major labelled components in mature bacteroids had estimated molecular weights of 60- and 34-kilodaltons similar to values reported by others for the constituent polypeptides of nitrogenase. Bacteroids of the same Rhizobium strain grown in different plant hosts gave similar polypeptide labelling patterns in purified suspensions, but bacteroids of different Rhizobium strains gave different patterns. The polypeptide labelling patterns obtained using broth-cultured Rhizobium bacteria from various growth stages and growth media differed from those obtained using bacteroids of the same strain.

Bacteria↗
PubMed1979

Carbon Dioxide Fixation by Lupin Root Nodules: II. Studies with C-labeled Glucose, the Pathway of Glucose Catabolism, and the Effects of Some Treatments That Inhibit Nitrogen Fixation.

Labeling studies using detached lupin (Lupinus angustifolius) nodules showed that over times of less than 3 minutes, label from [3,4-(14)C]glucose was incorporated into amino acids, predominantly aspartic acid, to a much greater extent than into organic acids. Only a slight preferential incorporation was observed with [1-(14)C]- and [6-(14)C]glucose, while with [U-(14)C]-glucose more label was incorporated into organic acids than into amino acids at all labeling times. These results are consistent with a scheme whereby the "carbon skeletons" for amino acid synthesis are provided by the phosphoenolpyruvate carboxylase reaction.A comparison of (14)CO(2) release from nodules supplied with [1-(14)C]- and [6-(14)C]glucose indicated that the oxidative pentose phosphate pathway accounted for less than 6% of glucose metabolism. Several enzymes of the oxidative pentose phosphate and glycolytic pathways were assayed in vitro using the 12,000g supernatant fraction from nodule homogenates. In all cases, the specific activities were adequate to account for the calculated in vivo fluxes.Three out of four diverse treatments that inhibited nodule nitrogen fixation also inhibited nodule CO(2) fixation, and in the case of the fourth treatment, replacement of N(2) with He, it was shown that the normal entry of label from exogenous (14)CO(2) into the nodule amino acid pool was strongly inhibited.

Journal Article↗
PubMed1977

Changes in the Number, Viability, and Amino-acid-incorporating Activity of Rhizobium Bacteroids during Lupin Nodule Development.

Between days 10 and 21 after inoculation of Lupinus angustifolius seedlings with Rhizobium NZP 2257, the average nodule fresh weight increased 3-fold and the number of bacteroids per nodule increased more than 10-fold.The viability of Rhizobium bacteroids, as judged by their ability to form colonies on yeast-extract agar, declined from about 10% on days 10 and 11 after inoculation to about 0.3% on days 14 to 25. Bacteroid viability was highly sensitive to osmolarity.At optimal pH and K and Mg ion concentrations, the incorporation of (14)C-glycine into isolated bacteroids was also very sensitive to osmolarity, and fell in parallel with bacteroid viability during nodule development.WE SUGGEST THAT AT LEAST TWO PROCESSES CONTRIBUTE TO BACTEROID NONVIABILITY: a reversible change in the cell wall structure occurring between days 10 and 14 after inoculation, and a subsequent irreversible change.

Journal Article↗
PubMed1977

Carbon Dioxide Fixation by Lupin Root Nodules: I. Characterization, Association with Phosphoenolpyruvate Carboxylase, and Correlation with Nitrogen Fixation during Nodule Development.

In vivo CO(2) fixation and in vitro phosphoenolpyruvate (PEP) carboxylase levels have been measured in lupin (Lupinus angustifolius L.) root nodules of various ages. Both activities were greater in nodule tissue than in either primary or secondary root tissue, and increased about 3-fold with the onset of N(2) fixation. PEP carboxylase activity was predominantly located in the bacteroid-containing zone of mature nodules, but purified bacteroids contained no activity. Partially purified PEP carboxylases from nodules, roots, and leaves were identical in a number of kinetic parameters. Both in vivo CO(2) fixation activity and in vitro PEP carboxylase activity were significantly correlated with nodule acetylene reduction activity during nodule development. The maximum rate of in vivo CO(2) fixation in mature nodules was 7.9 nmol hour(-1) mg fresh weight(-1), similar to rates of N(2) fixation and reported values for amino acid translocation.The results suggest that the oxaloacetete used as the primary "carbon skeleton" acceptor for ammonia assimilation and amino acid synthesis in lupin nodules is provided via the PEP carboxylase reaction rather than through the tricarboxylic acid cycle. The source of PEP is presumably glycolysis, while the major source of CO(2) is inferred to be respiration.

Journal Article↗
PubMed1977

Preparation and Fractionation of Rhizobium Bacteroids by Zone Sedimentation through Sucrose Gradients.

Zone sedimentation through sucrose gradients was used for preparing Rhizobium bacteroids from lupin nodules and for separating them into slowly and rapidly sedimenting fractions.The purity of the bacteroids was established by electron microscopy and by enzyme assays, and they were shown to contain a CO-insensitive cytochrome c oxidase.Bacteroids sedimented more rapidly than broth-cultured Rhizobium bacteria, and bacteroids from old nodules sedimented more rapidly than bacteroids from young nodules.THERE APPEAR TO BE AT LEAST TWO FORMS OF BACTEROID IN OLD NODULES: slowly sedimenting bacteroids with moderate colony-forming ability resembling the bacteroids found in young nodules, and rapidly sedimenting bacteroids with much lower colony-forming ability.

Journal Article↗
PubMed1975

Studies with detached lupin root nodules in culture: I. Maintenance and induction of acetylene reduction activity.

A method has been developed for culturing detached nitrogen-fixing root nodules of lupin (Lupinus angustifolius L.) on a simple nutrient medium. Under the best conditions devised, the acetylene reduction activity of mature detached nodules was maintained at 10 to 25 nmoles of ethylene hr(-1) mg(-1) fresh weight for 3 days. Under the same culture conditions, immature nodules increased their acetylene reduction activity from 0.01 nmole or less to about 1 nmole hr(-1) mg(-1) fresh weight.

Journal Article↗
PubMed1972

Self-renaturing fractions in the separated strands of mouse satellite deoxyribonucleic acid.

Pyrimidine- and purine-rich strands of Mus musculus satellite DNA prepared by alkaline CsCl-gradient centrifugation can self-renature to a variable extent to give partial duplexes with high thermal stability. These duplexes were purified by treatment with nuclease S(1) followed by hydroxylapatite chromatography, and have been shown by pyrimidine-tract analysis to be very similar in sequence to total reassociated satellite DNA. It is believed that the self-renaturing fractions result from variable contamination of each strand with fragments of the other, rather than from molecular inversions. The predominantly single-stranded properties of these fractions may be partly due to the ability of mouse satellite DNA strands to reassociate in non-stoicheiometric proportions.

Animals↗