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N Nanninga

Publications and source records attributed to N Nanninga.

At least 73 records · Page 4Linked to original sources
PubMed1980

Immunoferrin labeling of respiratory nitrate reductase in membrane vesicles of Bacillus licheniformis and Klebsiella aerogenes.

The indirect immunoferritin labeling method was used to localize the membrane-bound respiratory nitrate reductase in membrane vesicles and protoplasts or sphereplasts of Bacillus licheniformis and Klebsiella aerogenes, respectively. For a comparison of the labeling of the various vesicle preparations, which differed not only in size but also in the percentage of inside-out orientation, a quantification of the results was needed to circumvent the problem of non-specifically bound ferritin. From the results of sidedness of the nitrate reductase in the cytoplasmic membrane of the above-mentioned bacteria was determined as being cytoplasmic in B. licheniformis and as transmembranous in K. aerogenes.

Bacillus↗
PubMed1980

Positive correlation between size at initiation of chromosome replication in Escherichia coli and size at initiation of cell constriction.

The variability of (i) the length (size) at which cells initiate chromosome replication, (ii) the length at which they initiate cell constriction, and (iii) the time interval between these events has been estimated for Escherichia coli B/r K at two different slow growth rates. Steady-state cultures were pulse-labeled with [3H]thymidine and, after fixation, analyzed by electron microscopic radioautography. The coefficient of variation of length at initiation of chromosome replication was found to be 15 to 22%, the coefficient of variation of length at initiation of cell constriction was 10%, and the coefficient of variation of the time interval between both events was 25%. With the help of these values we calculated a high positive coefficient of correlation (rho) between the length at which a round of chromosome replication is initiated and that at which the onset of cell constriction occurs. At both growth rates rho has a value of 0.6 to 1.0. This correlation excludes a model in which chromosome initiation and cell constriction are independently triggered by some aspects of cell growth. It favors a model in which an event before or at chromosome initiation triggers both.

Cell Division↗
PubMed1980

Correlation between size and age at different events in the cell division cycle of Escherichia coli.

The variability of (i) the B period between birth and initiation of chromosome replication, (ii) the U period between initiation of chromosome replication and initiation of cell constriction, and (iii) the interdivision period (tau) have been estimated for slowly growing Escherichia coli B/r F. Cultures synchronized by the membrane elution technique were pulse-labeled with [3H]thymidine or continuously labeled with [3H]thymine. After fixation, the pattern of deoxyribonucleic acid replication was analyzed by electron microscopic radioautography. Cell length was found to increase exponentially with age at two different slow growth rates. The coefficient of variation of the B period was estimated to be 60%, that of the U period was 29%, and that of the interdivision period was 12%. From these values and the coefficient of variation of length at different cell cycle events were calculated a negative correlation between the B and U period (r = -0.9) and a positive correlation between length at birth and cell separation (r = 0.6). Initiation of chromosome replication and cell constriction were strictly correlated both with respect to age (r = 0.7) and length (r = 0.8). On the other hand, length at initiation of chromosome replication was distantly correlated with age (r = 0.1) or length at birth (r = 0.3). This low correlation excludes a model in which chromosome initiation is controlled by a random event in the B period. It favors a model in which chromosome initiation occurs at a particular distributed size independent of cell division.

Cell Cycle↗
PubMed1980

Pattern of meso-dl-2,6-diaminopimelic acid incorporation during the division cycle of Escherichia coli.

The topography of meso-DL-2,6-diaminopimelic acid incorportion into the cell envelope of Escherichia coli W7 (doubling time, tau, = 70 min) has been studied by autoradiography. To follow the incorporation pattern during the division cycle, cells have been classified according to length and the silver grain distributions have been determined in the two cell halves. In particular, the question of equivalence (with respect to the grain distributions) of the two cell halves has been investigated statistically. The grain localizations have been determined separately for the left cell halves (highest number of grains) and right cell halves. The highest probability of finding grains was in the central area for cells of all length classes. In the longest cells (dividing or nondividing) incorporation occurred in the future septal regions of the prospective daughter cells. Autoradiography of tritiated thymidine-labeled cells indicated the presence of an atypical deoxyribonucleic acid replication cycle (at tau = 70 min). Initiation of deoxyribonucleic acid replication occurred during the latter part of the division cycle, and its termination occurred in the next cycle.

Amino Acids, Diamino↗
PubMed1979

Formation of inside-out vesicles of Bacillus licheniformis. Dependence on buffer composition and lysis procedure.

1. The extent to which the cytoplasmic membrane of the Gram-positive bacterium Bacillus licheniformis formed inside-out vesicles was studied with the freeze-fracture technique. The membrane orientation appeared to be dependent on the buffer compositon as well as on the lysis procedure used. 2. By manipulating these conditions, membrane preparations were obtained with the percentage of inside-out vesicles varying from 15 to 80%. 3. More vesicles had the opposite orientation when the cells were lysed in potassium phosphate buffer than when they were lysed in sodium phosphate buffer. Tris-HCl buffer favoured the formation of inside-out vesicles more than phosphate buffer. 4. Lysis of protoplasts in hypotonic buffers resulted in more inside-out vesicles than did direct lysis of cells in hypotonic media. 5. In an attempt to explain the observed differences, experiments were performed in which the morphology of thin-sectioned lysing cells in sodium phosphate buffer was compared with that in potassium phosphate buffer. The results from these experiments indicate that the formation of inside-out vesicles is brought about by an effect on the membrane itself rather than on the cell wall, on the cell wall membrane association, or on the cytoplasm.

Bacillus↗
PubMed1979

Respiratory nitrate reductase: its localization in the cytoplasmic membrane of Klebsiella aerogenes and Bacillus licheniformis.

The sidedness of the respiratory nitrate reductase in the cytoplasmic membrane of Bacillus licheniformis and Klebsiella aerogenes was studied by indirect immunofluorescence and by lactoperoxidase-catalyzed iodination. It was shown that the two subunits (Mr 150000 and 57000, respectively) of nitrate reductase of B. licheniformis are localized on the cytoplasmic side of the membrane, whereas the K. aerogenes enzyme is a transmembrane protein. The different localization of nitrate reductase in the membranes of these organisms may be related to their different rôle in oxidative phosphorylation.

Bacillus↗
PubMed1979

The cell cycle of Bacillus subtilis as studied by electron microscopy.

Bacillus subtilis strain Marburg was grown exponentially with a doubling time of 65 min. To follow the time course of various cell cycle events, cells were collected by agar filtration and were then classified according to length. The DNA replication cycle was determined by a quantitative analysis of radioautograms of tritiated thymidine pulse labeled cells. The DNA replication period was found to be 45 min. This period is preceded and followed by periods without DNA synthesis of about 10 min. The morphology and segregation of nucleoplasmic bodies was studied in thin sections. B. subtilis contains two sets of genomes. DNA replication and DNA segregation seem to go hand in hand and DNA segregation is completed shortly after termination of DNA replication. Cell division and cell separation were investigated in whole mount preparations (agar filtration) and in thin sections. Cell division starts about 20 min after cell birth; cell separation starts at about 45 min and before completion of the septum.

Bacillus subtilis↗
PubMed1979

Length growth of two Escherichia coli B/r substrains.

Length growth of synchronized Escherichia coli B/r substrain A (ATCC 12407) and B/r substrain F26 (Thy his) was followed with an electron microscope. Cells were grown with doubling times (tau) of 60 min (B/rA) and of 82 and 165 min (B/rF26). Different length growth patterns were found for the two substrains. In B/rF, the length growth rate increased about midway in the cell cycle. For tau = 165 min, the rate increase was preceded by a short period of slow growth. For B/r A (r = 60 min), this period seemed to occur at the beginning of the cell cycle. The possibility is raised that the different length growth patterns are related to different deoxyribonucleic acid replication patterns of the respective strains.

Cell Cycle↗
PubMed1978

DNA replication pattern and cell wall growth in Escherichia coli PAT 84.

An electron microscopic radioautographic study was made of tritiated thymidine incorporation into the genome of Escherichia coli PAT 84 and of tritiated meso-D,L-2,6-diaminopimelic acid (DAP) into the cell envelope. Pulse-labeled cells growing at 30 degrees C with a doubling time of 170 min were classified according to length by the method of agar filtration. Mathematical analysis of the length distribution led to the assumption of an exponential relation between length and time. A novel DNA replication pattern was found. Within the cell cycle DNA replication terminates at 70 min; then a gap follows of 64 min, after which DNA replication is initiated at 134 min. Thus, the C period is 106 min and the D period is 100 min. Cell constriction starts at 141 min and coincides with initiation of DNA replication. Detailed quantitative analysis of the [3H]thymidine grain frequency distribution allowed the distinction of three groups of cells. The first group incorporated no label, the second group an amount C, and the third group an amount 2 X C. The relative contribution of each group to a particular length class was determined. The data fitted very well into the DNA replication pattern. The same analysis was carried out on DAP pulse-labeled cells. Again, three groups of cells could be distinguished, and their relative contributions to each length class was determined. The group with the double amount of label was especially prominent at the end of the cell cycle. The emergence of this group might represent the acquisition of new lateral growth areas.

Cell Cycle↗
PubMed1978

Size variations and correlation of different cell cycle events in slow-growing Escherichia coli.

Cell lengths have been determined at which cycle events occur in the slow-growing Escherichia coli B/r substrains A, K, and F26. The radioautographic and electron microscope analyses allowed determination of the variations in length at birth, initiation and termination of DNA replication, and initiation of the constriction process and of cell separation. In all three substrains the standard deviation increased between cell birth and initiation of DNA replication. From there on, the standard deviation remained relatively constant until cell separation. These observations are consistent with the presence of a deterministic phase during the cell cycle in which the cell sizes at initation of DNA replication and at cell division are correlated.

Cell Cycle↗
PubMed1978

Arrangement of glycan chains in the sacculus of Escherichia coli.

A novel of Escherichia coli endopeptidase was used for a selective partial hydrolysis of the peptide bridges which interlink the glycan chains in E. coli sacculi. The loosening of the murein network revealed, in the electron microscope, a preferential orientation of the glycan chains, more or less perpendicular to the length axis of the cell. Control incubations with E. coli transglycosylase or egg-white lysozyme did not leave ordered structures behind.

Escherichia coli↗
PubMed1976

An investigation of the binding sites of proteins S8, L23 and L24 on the ribosomal RNAs of Escherichia coli by electron microscopy.

An electron microscopic method was used to investigate the binding regions of proteins S8 on 16S RNA, and proteins L23 and L24 on 23S RNA. Regions of the RNA that were not stabilised by the protein were completely denatured in 80% dimethyl-sulfoxide. The lengths of these denatured RNA regions were compared with that of the whole denatured RNA. Conclusions are drawn concerning the approximate location of the three proteins and these results are correlated with both RNA structural data and RNA sequence data on the RNA binding regions of the proteins.

Bacterial Proteins↗
PubMed1976

Significance of folded chromosomes released from amino-acid-starved Escherichia coli cells.

1. Folded chromosomes from amino-acid-starved Escherichia coli DG 75 cells are to a large extent released as envelope-bound complexes which sediment more rapidly than envelope-bound complexes from exponentially growth cells. A minor fraction (about 3%) represents relatively slow sedimenting envelope-free nucleoids. 2. Morphological analysis of the sensitivity of amino-acid-starved cells to the action of lysozyme and/or detergents indicates that these cells are less susceptible to lysis than exponentially grown cells. This results in the production of fast sedimenting envelope-bound complexes from non-dividing cells. We infer that it is not the amount of DNA, as suggested by Ryder and Smith (1974), but the low degree of envelope fragmentation that causes the high sedimentation rate. 3. After prolonged periods of starvation about 3% of cells in the process of division persist in the population. The results indicate that these cells release their (terminated) chromosomes in the envelope-free form. At this stage it is impossible to conclude whether these chromosomes are released because of their detachment from the membrane in situ (cf. Worcel and Burgi, 1974) or because of an enhanced susceptibility of dividing cells to lysis.

Amino Acids↗
PubMed1976

Factors affecting the release of folded chromosomes from Escherichia coli.

1. The envelope-bound folded chromosome prepared by gentle lysis of Escherichia coli K 12 DG 75 cells in the presence of 1.0 M sodium chloride was found to be a non-specific complex of DNA strands entangled in vesicle-like envelope structures. This envelope remnant is still more or less rod-shaped. 2. Both the envelope-bound and the envelope-free folded chromosomes arise directly from intact cells. Even under extreme conditions of lysis, envelope-bound chromosomes cannot be converted into envelope-free nucleoids.

Cell Fractionation↗
PubMed1976

Localization of ampicillin-sensitive sites in Escherichia coli by electron microscopy.

Growth of Escherichia coli B/r ATCC 12407 (doubling time, 65 to 70 min) in the presence of 500 mug of ampicillin per ml for 15 to 20 min induces a sphere alongside the cell. The position was determined with respect to the length axis of the cell by electron microscopy. Although spheres may be found anywhere, some prominent sites do occur. In the shortest cells, which have a length of about 1.5 mum, they are found at the presumed new cell pole. In slightly older cells (length, about 1.8 mum), the position of the sphere is not well defined. Later on spheres occur predominantly at the cell center. In dividing cells (average length, 2.5 mum) a sphere may also occur at about one-quarter of the cell length. The position of the spheres bears resemblance to sites where a pulse of 3H-labeled diaminopimelic acid is incorporated into the peptidoglycan, as has been found by others.

Ampicillin↗
PubMed1976

Organization of the nucleoplasm in Escherichia coli visualized by phase-contrast light microscopy, freeze fracturing, and thin sectioning.

The organization of the nucleoplasm in Escherichia coli was studied by comparing the results obtained by freeze fracturing and thin sectioning. In addition to exponentially growing cells, we used chloramphenicol-treated cells which show a well-defined nucleoplasm, in the phase-contrast light microscope and can therefore function as a control for treatments necessary for electron microscopy. Two factors were found to determine the visibility of the nucleoplasm in freeze fractures: first, the state of lateral aggregation of deoxyribonucleic and fibrils, which is enhanced by postfixation with OsO4 according to the Ryter-Kellenberger technique; second, the presence of ice crystals. When their formation is prevented by the use of high concentration of freeze-protecting agents, the nucleoplasm appears as a smooth region in cells that have been prefixed. In unfixed cells, however, the freeze-protecting agent causes disappearance of the nucleoplasm by rearrangement of structures within the cell. This observation makes it hard to determine whether the deoxyribonucleic acid in vivo dispersed, as found after glutaraldehyde prefixation, or compact, as after OsO4 prefixation.

Cell Nucleus↗