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Biomedical subjects

G Turnock

Publications and source records attributed to G Turnock.

At least 19 recordsLinked to original sources
PubMed1996

The Escherichia coli enzoskeleton.

The nature of the structure of the bacterial cell is becoming clearer. The envelope contains periseptal annuli, a discontinuous periplasm and adhesion sites, whilst the cytoplasmic membrane is probably organized into distinct proteolipid domains by the coupled transcription-translation-insertion (transertion) of membrane proteins. The structure of the nucleoid is determined by proteins which self-associate and by attachment to membrane, which is achieved in part by transertion. Metabolic pathways form multi-enzyme complexes which channel substrates and which connect membranes and nucleic acids to create the extensive, cross-linked, intracellular structure we term the 'enzoskeleton'. This enzoskeleton includes eukaryotic-like cytoskeletal structures and elements such as the MukB and FtsZ proteins. We propose that the enzoskeleton is regulated by calcium and by protein phosphorylation during adaptation to different environments and during the cell cycle.

Actins↗
PubMed1987

Differential gene expression during the amoebal-plasmodial transition in Physarum.

We have prepared cDNA libraries for amoebae and plasmodia of the acellular slime mould, Physarum polycephalum. Differential screening was used to isolate cell-type-specific cDNA clones (in bacteriophage M13) and both libraries yielded approximately 5% of such sequences. The amoebal- and plasmodial-specific clones were used to assay changes in transcription during the amoebal-plasmodial transition. The results obtained substantiate the view that the switch from amoebal to plasmodial characteristics occurs over several nuclear division cycles. With one exception, the specific cDNAs came from single-gene families. Southern blotting experiments also showed that they hybridised to identical restriction fragments from amoebal and plasmodial DNAs indicating that genomic rearrangements are unlikely to be involved in the regulation of these genes.

Cloning, Molecular↗
PubMed1985

In vivo transcription of ribosomal RNA in relation to the mitotic cycle in Physarum polycephalum.

We have investigated the transcription of ribosomal RNA in plasmodia of Physarum polycephalum by a combination of pulse-labelling with [3H]uridine and RNA:DNA hybridization. The DNA used for the hybridization was a HindIII restriction fragment (cloned in bacteriophage lambda) of Physarum ribosomal DNA that carries a substantial fraction of the rRNA genes, enabling the ribosomal transcripts in the newly synthesized RNA to be measured. We found that ribosomal RNA constituted about 40% of the pulse-labelled RNA at all times during the synchronous mitotic cycle.

DNA↗
PubMed1982

Transfer RNA and ribosomal RNA are synthesized from the same pyrimidine nucleotide pool.

We have tested the hypothesis [Wiegers, Kramer, Klapproth & Hilz (1976) Eur. J. Biochem. 64. 535-540] that the synthesis of rRNA in the nucleolus may have a partially independent supply of nucleoside triphosphates that is not completely suppressed when cells are supplied with exogenous nucleosides. For the slime mould, Physarum polycephalum, and Chinese hamster ovary cells in culture, the specific activities (3H/32P) of UMP and CMP in tRNA, which is synthesised in the nucleoplasm, and rRNA were compared after continuous labelling with [3H]uridine and [32P]phosphate. No differences were found, suggesting that transcription throughout the nucleus draws on a common supply of pyrimidine triphosphates that is uniformly labelled from exogenous [3H]uridine. As a control of the radioactive labelling schedule, a similar experiment was carried out with the prokaryote, Escherichia coli, and identical results were obtained.

Animals↗
PubMed1981

Regulation of protein synthesis in the plasmodial phase of Physarum polycephalum.

1. The rate of protein synthesis changes very little during the first 2-3 h (S phase) of the nuclear division cycle in plasmodia of Physarum polycephalum and then increases continuously during G2 phase, so that by the end of the cycle the rate has doubled relative to that in S phase. Protein synthesis appears to continue during mitosis. 2. Fractionation of extracts of plasmodia, labeled with [3H]lysine for 1 h, by two-dimensional electrophoresis indicated that most if not all proteins are synthesised throughout the nuclear division cycle. However, two metabolically stable polypeptides, the synthesis of which occurs predominantly in Gs phase, were detected. 3. Using a double-labelling procedure, the differential rates of synthesis of 30 relatively abundant polypeptides were measured in relation to the nuclear division cycle. As a group, their differential rates of synthesis increase during the cycle so that their actual rates of synthesis increase 4-6-fold. This implies that their synthesis is regulated over and above any simple change due to a doubling in the number of genes during S phase.

Cell Cycle↗
PubMed1981

A comparison of the proteins of the amoebal and plasmodial phases of the slime mould, Physarum polycephalum.

1. The proteins of amoebae and plasmodia of strain CL of Physarum polycephalum have been compared by two-dimensional gel electrophoresis. Both forms of the organism were labelled by growth on formalin-killed bacteria labelled with [35S]sulphate, [3H]lysine or [14C]lysine. Plasmodia were also labelled from radioactive lysine in the medium. 2. Of 306 relatively abundant proteins examined, 26% were phase-specific, that is they were found only in amoebae or in plasmodia. About a quarter of these apparent differences in gene expression may be due to minor changes in charge and/or size. 3. Amongst the 74% of the proteins present in both amoebae and plasmodia, there are substantial differences in differential rates of synthesis and these have been measured for a representative set of proteins by a double-label procedure.

Carbon Radioisotopes↗
PubMed1977

Synthesis of transfer RNA during the synchronous nuclear division cycle in Physarum polycephalum.

1. The synthesis of tRNA during the synchronous nuclear division cycle in plasmodia of Physarum polycephalum has been investigated using an isotope dilution procedure and compared to the pattern of synthesis of rRNA. 2. The synthesis of both tRNA stops during mitosis. As the genes for the two types of RNA are not linked, this finding suggests that cessation of synthesis during nuclear division is a common characteristic of all types of transcription, supporting earlier work using pulse labelling and autoradiography to investigate the same problem.

Cell Division↗
PubMed1976

The uptake and metabolism of uridine by the slime mould Physarum polycephalum.

1. Uridine is taken up by microplasmodia of Physarum polycephalum via a saturatable transport system with an apparent Km of 29 muM. An intracellular concentration significantly higher than that in the growth medium is attained, suggesting that the uptake is an active process. Both deoxyribonucleosides and ribonucleosides are competitive inhibitors of the uptake of uridine. 2. In contrast, the rate of entry of uridine into surface plasmodia is a linear function of the concentration of the nucleoside in the growth medium, and the uptake is not inhibited by other nucleosides. 3. As well as serving as a source of pyrimidine nucleotides for the synthesis of nucleic acids, uridine is also catabolised by P. polycephalum. Uracil accumulates in the growth medium and there is also significant conversion of C-2 of the pyrimidine ring to CO2. The proportion of uridine subject to catabolism in surface plasmodia is less than that observed for microplasmodia.

Binding, Competitive↗
PubMed1976

Synthesis of ribosomal RNA during the mitotic cycle on the slime mould Physarum polycephalum.

1. An isotope dilution technique has been used to analyze the synthesis of metabolically stable nucleic acids during the mitotic cycle in surface plasmodia of the slime mould Physarum polycephalum. Microplasmodia that had been labelled with [3H]uridine were used to prepare a surface culture, after a period of growth long enough to ensure that radioactivity was present only in tRNA, rRNA and DNA. The synthesis of rRNA or nuclear DNA during the growth of the surface plasmodium was then followed by measuring the specific activity of the nucleic acid. 2. Synthesis of rRNA during the mitotic cycle shows the following characteristics: (a) it is low during the immediate period of nuclear division, (b) synthesis is then continuous throughout interphase and (c)the rate of synthesis increases 5-6-fold between the beginning and end of interphase. These results are discussed in relation to the pattern of replication of the genes for rRNA. 3. Approximately 80% of the nuclear DNA replicates during the first 90 min of the mitotic cycle; completion of replication, however, occupies the remainder of interphase.

Cell Nucleus↗
PubMed1975

Ribosomal RNA genes in the amoebal and plasmodial forms of the slime mould Physarum polycephalum.

1. The degree of homology between ribosomal RNA isolated from microplasmodia and amoebae of the slime mould Physarum polycephalum has been determined by competive hybridisation of the RNA from the two sources to homologous DNA in solution. The extent of competition was measured both by hybridisation to saturation and by following the kinetics of hybrid formation. In each case competition was found to be 100%, indicating that the ribosomal RNAs from the two, quite different vegetative forms of the organism exhibit a high degree of homology and are probably transcribed from the same genes. 2. The relationship between the amount of nuclear DNA that codes for ribosomal RNA (rDNA) and ploidy has been investigated in three strains of P. polycephalum which exhibit a 1:2:5 variation in the amount of DNA per nucleus. Ribosomal RNA saturation values were determined by hybridisation to DNA isolated from prophase nuclei of plasmodia. The proportion of rDNA was found to be constant at 0.16--0.18% of the total genome in the three strains.

Amoeba↗
PubMed1975

Coordination of macromolecular synthesis in the slime mould Physarum polycephalum.

Microplasmodia of P. polycephalum were grown either in batch culture, in both complex and defined media to give a 3-4 fold variation in growth rate, or in a chemostate. The protein/DNA ratio of batch cultures was almost invariant, whilst the RNA/DNA ratio increased as a non-linear function of growth rate. The amount of ribosomal RNA, expressed as a fraction of total RNA, showed little variation and this was also true for the proportion of ribosomes found in polyribosomes. Calculation of the rate of protein synthesis per ribosome shows that this parameter increases by approximately 50% over the range of growth rates studied, although it should be emphasized that the effect of protein turnover has not yet been taken into account. Enrichment of batch cultures growing in a defined medium produced an increase in the rate of RNA synthesis. Data obtained with chemostat cultures differed in several respects from those described above for batch cultures, especially at low growth rates, and are discussed in relation to the early stages of differentiation of microplasmodia to spherules.

DNA↗