Showing papers in "Journal of Bacteriology in 1970"
TL;DR: In bioluminescent bacteria growing in shake flasks, the enzyme luciferase has been shown to be synthesized in a relatively short burst during the period of exponential growth, attributed to a stimulation of existing patterns of synthesis.
Abstract: In bioluminescent bacteria growing in shake flasks, the enzyme luciferase has been shown to be synthesized in a relatively short burst during the period of exponential growth. The luciferase gene appears to be completely inactive in a freshly inoculated culture; the pulse of preferential luciferase synthesis which occurs later is the consequence of its activation at the level of deoxyribonucleic acid transcription which is attributed to an effect of a “conditioning” of the medium by the growing of cells. Although cells grown in a minimal medium also exhibit a similar burst of synthesis of the luminescent system, the amount of synthesis is quantitatively less, relative to cell mass. Under such conditions, added arginine results in a striking stimulation of bioluminescence. This is attributed to a stimulation of existing patterns of synthesis and not to induction or derepression per se.
1,174 citations
TL;DR: In this paper, the base composition of deoxyribonucleic acid (DNA) was calculated by regression and correlation analysis and treated statistically by using only sets of data on DNA determined with the same strains.
Abstract: The equations currently used for the calculation of the chemical base composition of deoxyribonucleic acid (DNA), expressed as moles per cent guanine plus cytosine (% GC), from either buoyant density (ρ) or midpoint of thermal denaturation (Tm) were recalculated by using only sets of data on DNA determined with the same strains. All available information from the literature was screened and supplemented by unpublished data. The results were calculated by regression and correlation analysis and treated statistically. From the data on 96 strains of bacteria, it was calculated that% GC = 2.44 (Tm – 69.4). Tm appears to be unaffected by the substitution of cytosine by hydroxymethylcytosine. This equation is also valid for nonbacterial DNA. From the data on 84 strains of bacteria, the relation% GC = 1038.47 (–1.6616) was calculated. The constants in this equation are slightly modified when data on nonbacterial DNA are included. Both correlations differ only slightly from those currently used, but now they lean on a statistically sound basis. As a control, the relation between ρ and Tm was calculated from data of 197 strains; it agrees excellently with the above two equations.
569 citations
TL;DR: The findings are consistent with the hypothesis that vesicles produced by the endomembrane system in the subapical region become concentrated in the apex where they are incorporated at the expanding surface.
Abstract: Hyphal tips of fungi representing Oomycetes, Zygomycetes, Ascomycetes, Basidiomycetes, and Deuteromycetes were examined by light and electron microscopy and compared with respect to their protoplasmic organization. In all fungi studied, there is a zone at the hyphal apex which is rich in cytoplasmic vesicles but nearly devoid of other cell components. Some vesicle profiles are continuous with the plasma membrane at the apices of these tip-growing cells. The subapical zones of hyphae contain an endomembrane system which includes smooth-surfaced cisternae associated with small clusters of vesicles. The findings are consistent with the hypothesis that vesicles produced by the endomembrane system in the subapical region become concentrated in the apex where they are incorporated at the expanding surface. Septate fungi (Ascomycetes, Basidiomycetes, and Deuteromycetes) have an apical body (Spitzenkorper) which is associated with growing hyphal tips. In electron micrographs of these fungi, an additional specialized region within the accumulation of apical vesicles is shown for the first time. This region corresponds on the bases of distribution among fungi, location in hyphae, size, shape and boundary characteristics to the Spitzenkorper seen by light microscopy. This structure is not universally associated with tip growth, whereas apical vesicles are widespread among tip-growing systems.
429 citations
TL;DR: The composition of a defined nongrowth medium used in stage II development of competence of Haemophilus influenzae affects the course of this development and the method of evaluating competence, which depends on the frequency of multiple independent transformations, has been reexamined.
Abstract: The composition of a defined nongrowth medium used in stage II development of competence of Haemophilus influenzae affects the course of this development. The development of competence in two nongrowth media, M-IV and M-V, is rapid, logarithmic, and independent of the cell concentration. This last property indicates that there is probably no transfer of a competence factor from competent to noncompetent cells, in contrast to results reported for other organisms. Levels of competence reached in these completely defined media are such that 1 to 5% of the cells are transformed in the presence of an excess of marked deoxyribonucleic acid. The method of evaluating competence, which depends on the frequency of multiple independent transformations, has been reexamined. This and other methods are compared on samples taken from a culture during development of competence.
405 citations
TL;DR: Envelope preparations obtained by passing Escherichia coli cells through a French pressure cell were separated by sucrose density gradient centrifugation into two distinct particulate fractions, and one of the proteins which is clearly localized in the cell wall is the protein with a molecular weight of 44,000, which is the major component of the envelope.
Abstract: Envelope preparations obtained by passing Escherichia coli cells through a French pressure cell were separated by sucrose density gradient centrifugation into two distinct particulate fractions. The fraction with the higher density was enriched in fragments derived from the cell wall, as indicated by the high content of lipopolysaccharide, the low content of cytochromes, and the similar morphology of the fragments and intact cell walls. The less-dense fraction was enriched in vesicles derived from the cytoplasmic membrane, as indicated by the enrichment of cytochromes, the enzymes lactic and succinic dehydrogenase and nitrate reductase, and the morphological similarity of the vesicles to intact cytoplasmic membrane. Both fractions were rich in phospholipid. The protein composition was compared by mixing the cytoplasmic membrane-enriched fraction from a (3)H-labeled culture with the cell wall-enriched fraction from a (14)C-labeled culture and examining the resulting mixture by gel electrophoresis. Thirty-four bands of radioactive protein were resolved; of these, 27 were increased two- to fourfold in the cytoplasmic membrane-enriched fraction, whereas 6 were similarly increased in the cell wall-enriched fraction. One of the proteins which is clearly localized in the cell wall is the protein with a molecular weight of 44,000, which is the major component of the envelope. This protein accounted for 70% of the total protein of the cell wall, and its occurrence in the envelope from spheroplasts suggests that it is a structural protein of the outer membranous component of the cell wall.
401 citations
TL;DR: Kinetic and genetic evidences are presented to show that, in addition to specific amino acid permeases, Saccharomyces cerevisiae has a general amino acids permease which catalyzes the transport of basic and neutral amino acids, but most probably not that of proline.
Abstract: Kinetic and genetic evidences are presented to show that, in addition to specific amino acid permeases, Saccharomyces cerevisiae has a general amino acid permease which catalyzes the transport of basic and neutral amino acids, but most probably not that of proline. The general amino acid permease appears to be constitutive, and its activity is inhibited when ammonium ions are added to the culture medium. A mutant which has lost the general amino acid permease activity was isolated. Its mutation, named gap (general amino acid permease), is not allelic to the aap (amino acid permease) mutation of Surdin et al., which has a quite different phenotype and cannot be considered as having selectively lost the general amino acid permease activity.
335 citations
TL;DR: Samples of bacterial deoxyribonucleic acid from bacteria having guanine plus cytosine (GC) contents in the range of 27 to 72 moles per cent GC were analyzed by optical melting (T(m)) and equilibrium buoyant density methods and the relation between these properties is shown to be linear.
Abstract: Samples of bacterial deoxyribonucleic acid from bacteria having guanine plus cytosine (GC) contents in the range of 27 to 72 moles per cent GC were analyzed by optical melting (T(m)) and equilibrium buoyant density methods. The relation between these properties is shown to be linear. The relative value of 1.99 moles per cent GC per degree C change in T(m) is calculated, and a reference method for the calculation of GC contents relative to a standard is derived.
309 citations
TL;DR: Intra- and intergroup relationships obtained from the numerical taxonomy studies showed highly significant correlation with DNA/DNA reassociation data.
Abstract: A set of 86 bacterial cultures, including 30 strains of Vibrio cholerae, 35 strains of V. parahaemolyticus, and 21 representative strains of Pseudomonas, Spirillum, Achromobacter, Arthrobacter, and marine Vibrio species were tested for a total of 200 characteristics. Morphological, physiological, and biochemical characteristics were included in the analysis. Overall deoxyribonucleic acid (DNA) base compositions and ultrastructure, under the electron microscope, were also examined. The taxonomic data were analyzed by computer by using numerical taxonomy programs designed to sort and cluster strains related phenetically. The V. cholerae strains formed an homogeneous cluster, sharing overall S values of >/=75%. Two strains, V. cholerae NCTC 30 and NCTC 8042, did not fall into the V. cholerae species group when tested by the hypothetical median organism calculation. No separation of "classic" V. cholerae, El Tor vibrios, and nonagglutinable vibrios was observed. These all fell into a single, relatively homogeneous, V. cholerae species cluster. V. parahaemolyticus strains, excepting 5144, 5146, and 5162, designated members of the species V. alginolyticus, clustered at S >/=80%. Characteristics uniformly present in all the Vibrio species examined are given, as are also characteristics and frequency of occurrence for V. cholerae and V. parahaemolyticus. The clusters formed in the numerical taxonomy analyses revealed similar overall DNA base compositions, with the range for the Vibrio species of 40 to 48% guanine plus cytosine. Generic level of relationship of V. cholerae and V. parahaemolyticus is considered dubious. Intra- and intergroup relationships obtained from the numerical taxonomy studies showed highly significant correlation with DNA/DNA reassociation data.
307 citations
TL;DR: The taxonomic significance of production of hydroxamic acids is described in connection with the position of these yeast species in the subclass Heterobasidiomycetidae.
Abstract: An examination of 142 strains within 19 genera of yeasts and yeastlike organisms for formation of hydroxamic acids in low-iron culture showed production of hydroxamates by two unclassified strains and by 52 strains among the genera Aessosporon (3 of 3 strains), Cryptococcus (1 of 43), Leucosporidium (3 of 11), Rhodosporidium (4 of 4), Rhodotorula (27 of 39), Sporidiobolus (2 of 2), and Sporobolomyces (12 of 13). Crystalline rhodotorulic acid was isolated in amounts sufficient to account for most or all of the measured hydroxamate in culture supernatants of 16 strains representative of the five last-mentioned hydroxamate-producing genera. A new alanine-containing ferrichrome was isolated from one strain of Cryptococcus melibiosum. Rhodotorulic acid was a major metabolic product of many of the positive strains when grown in low-iron media, and iron was shown to repress its synthesis and excretion into the culture medium. The taxonomic significance of production of hydroxamic acids is described in connection with the position of these yeast species in the subclass Heterobasidiomycetidae.
282 citations
TL;DR: An endonuclease purified approximately 3,200-fold from Micrococcus luteus has all the attributes hypothesized for the first enzyme in the sequential steps in repair of DNA in vivo and is able to reactivate ultraviolet-irradiated transforming DNA.
Abstract: An endonuclease purified approximately 3,200-fold from Micrococcus luteus is active on native ultraviolet-irradiated deoxyribonucleic acid (DNA), but is inactive on unirradiated native or denatured DNA and has no activity toward irradiated denatured DNA. The major type of lesion for the nucleolytic activity is the cyclobutane pyrimidine dimer. The enzyme makes a number of single-strand breaks approximately equal to the number of dimers, but dimers are not excised. This endonuclease-a small molecular weight protein-therefore has all the attributes hypothesized for the first enzyme in the sequential steps in repair of DNA in vivo. Another paper shows that the endonuclease is able to reactivate ultraviolet-irradiated transforming DNA.
275 citations
TL;DR: Two infectious drug-resistance (R) factors, R28K and R6K, each conferring resistance to a number of penicillins by the synthesis of a penicillinase, were transferred to Proteus mirabilis PM1 and Escherichia coli RC85 host strains and showed correlation between the estimated number of R-factor copies present as CCC molecules and the enzyme activity per cell.
Abstract: Two infectious drug-resistance (R) factors, R28K and R6K, each conferring resistance to a number of penicillins by the synthesis of a penicillinase, were transferred to Proteus mirabilis PM1 and Escherichia coli RC85 host strains. Deoxyribonucleic acid (DNA) extracted from these strains was separated by density-gradient centrifugation and subjected to electron microscopy by use of a modification of the protein-monolayer diffusion technique. Analytical density-gradient centrifugation of the purified DNA from PM1 strains showed, in addition to the major peak at a density of 1.698 g/cm(3) characteristic of Proteus chromosomal DNA, a single satellite band at a density of 1.710 g/cm(3) [guanine plus cytosine (GC) base ratio 50%] for R28K and at 1.704 g/cm(3) (GC base ratio 45%) for R6K. Direct CsCl density-gradient centrifugation of crude lysates of the E. coli (R28K)(+) strain in the presence of ethidium bromide gave rise to a sedimentation profile with a single satellite peak containing covalently closed circular (CCC) DNA molecules with a mean contour length of 21.4 mum [44 x 10(6) atomic mass units (AMU)], although a minority was 13.6 mum in length. From the size of the major class, it was estimated that there were two to three copies of the R28K factor present as CCC molecules per chromosome at various phases of cellular growth. Similar studies of the E. coli (R6K)(+) lysates showed two satellite peaks; peak I contained mostly CCC molecules of contour length 12.8 mum (26 x 10(6) AMU), and peak II, intermediate to peak I and the chromosomal peak, contained CCC molecules of a similar size, together with about equal numbers of catenated molecules, mostly dimers consisting of two interlocked monomers of 12.8 mum. A smaller number (ca. 0.1%) of higher catenanes was also seen. The number of CCC copies of the R6K factor per chromosome present in peak I was calculated as 13 at logarithmic phase and 38 at stationary phase. In peak II, a constant ratio of about one catenated dimer per chromosome was found at all phases of growth. Penicillinase assays of cultures at different phases of growth showed a correlation between the estimated number of R-factor copies present as CCC molecules and the enzyme activity per cell for both R28K and R6K.
TL;DR: It is concluded that penicillin inhibits binary fission and prevents the synthesis of certain components essential for the formation of the elementary body envelope.
Abstract: L cells were infected at high multiplicity with meningopneumonitis organisms and incubated in medium containing 200 units per ml of penicillin. At intervals up to 48 hr after infection, cells were removed and thin sections were prepared for electron microscopic studies on the morphology of the developing organism. Penicillin had no effect on the initial reorganization of the infecting elementary body to form the developmental reticulate body (RB), and, up to 12 hr after infection, the treated and untreated cultures were identical. After that time, however, penicillin-treated organisms showed striking differences in that binary fission was prevented, large abnormal RB forms were seen in great numbers, masses of RB cytoplasmic membranes and envelopes were formed within and outside the RB itself, and large numbers of empty or partially filled small vesicles were pinched off the RB. After 36 hr immature nucleoids were formed within the RB. Throughout all of this period, both the outer cell envelope and the cytoplasmic membrane of these RB were recognized. When infected cells were transferred into penicillin-free medium, the abnormal RB showed recovery to form normal RB both by a budding-like process and by internal fragmentation or subdivision rather like endosporulation. We have concluded that penicillin inhibits binary fission and prevents the synthesis of certain components essential for the formation of the elementary body envelope.
TL;DR: A motile flagellate ( fla(+)) variant of a fim(-)fla(-) strain of S. typhimurium outgrew its parent strain in mixed cultures in aerobic static broth, but the selective advantage conferred by motility was weaker than that conferred by fimbriation.
Abstract: Competitive mixed cultures were grown from inocula of a large number of bacteria of a genotypically nonfimbriate (fim−) strain of Salmonella typhimurium and a small number of a genotypically fimbriate (fim+) variant strain that formed type 1 fimbriae and had been derived from the fim− strain by phage transduction. The fim+ strain differed from the fim− strain in fermenting l-rhamnose (rha+), and the viable fim+ and fim− bacteria present in the cultures after different periods at 37 C were counted differentially in platings on rhamnose media. When the cultures were grown under aerobic static conditions in tubes of nutrient broth, the fim+ bacteria rapidly outgrew the fim− bacteria, so that, although starting as a small minority (e.g., 1 in 107), they approached or surpassed the number of the fim− in 48 hr. A pellicle consisting of fimbriate bacteria was formed on the surface of the broth between 6 and 24 hr, and it is thought that the advantage of access to atmospheric oxygen enjoyed by these bacteria in the pellicle enabled them to outgrow the fim− bacteria confined in the oxygen-depleted broth. The fim+ bacteria did not show selective outgrowth in mixed cultures grown in broth aerated by continuous shaking, in static broth incubated anaerobically in hydrogen, and on aerobic agar plates, i.e., under conditions not allowing an advantage from pellicle formation. The outgrowth of fim+ bacteria in aerobic static broth was prevented by the addition of α-methylmannoside, a substance that inhibits the adhesive and early pellicle-forming properties of bacteria with type 1 fimbriae. A motile flagellate (fla+) variant of a fim−fla− strain of S. typhimurium outgrew its parent strain in mixed cultures in aerobic static broth, but the selective advantage conferred by motility was weaker than that conferred by fimbriation.
TL;DR: It is concluded that nonmethanogenic microbes metabolize intercellular formate in the rumen, and CO(2) and H( 2) are the principal substrates for rumen methanogenesis.
Abstract: An average of 11 (range, 2 to 47) mumoles of formate per g per hr was produced and used in whole bovine rumen contents incubated in vitro, as calculated from the product of the specific turnover rate constant, k, times the concentration of intercellular formate. The latter varied between 5 and 26 (average, 12) nmoles/g. The concentration of formate in the total rumen contents was as much as 1,000 times greater, presumably owing to formate within the microbial cells. The concentration of formate in rumen contents minus most of the plant solids was varied, and from the rates of methanogenesis the Michaelis constant, K(m), for formate conversion to CH(4) was estimated at 30 nmoles/g. Also, the dissolved H(2) was measured in relation to methane production, and a K(m) of 1 nmole/g was obtained. A pure culture of Methanobacterium ruminantium showed a K(m) of 1 nmole of H(2)/g, but the K(m) for formate was much higher than the 30 nmoles for the rumen contents. It is concluded that nonmethanogenic microbes metabolize intercellular formate in the rumen. CO(2) and H(2) are the principal substrates for rumen methanogenesis. Eighteen per cent of the rumen methane is derived from formate, as calculated from the intercellular concentration of hydrogen and formate in the rumen, the Michaelis constants for conversion of these substrates by rumen liquid, and the relative capacities of whole rumen contents to ferment these substrates.
TL;DR: Adenosine-3',5'-cyclic phosphate (cyclic AMP) is absolutely required for flagella formation and, hence, motility in cyclicAMP-deficient mutants of Escherichia coli and Salmonella typhimurium.
Abstract: Adenosine-3′,5′-cyclic phosphate (cyclic AMP) is absolutely required for flagella formation and, hence, motility in cyclic AMP-deficient mutants of Escherichia coli and Salmonella typhimurium.
TL;DR: Mutants of Diplococcus pneumoniae that lacked the two major deoxyribonucleases of the cell were obtained, indicating that a particular cellular component that is susceptible to loss by mutation, such as an enzyme, is responsible for low integration efficiency.
Abstract: Mutants of Diplococcus pneumoniae that lacked the two major deoxyribonucleases of the cell—one an endonuclease, the other an exonuclease preferentially active on native deoxyribonucleic acid (DNA)—were obtained. The development of a method for detecting mutant colonies, based on the binding of methyl green to DNA, facilitated isolation of the mutants. Neither enzyme was essential for growth of the cells, for repair of ultraviolet damage, or for any phase of DNA-mediated transformation. Residual deoxyribonuclease activity in the double mutant corresponded to an exonuclease, approximately one-fifth as active as the major exonuclease, that attacked native and denatured DNA equally well. This activity appeared to be associated with the DNA-polymerase enzyme. A mutant that apparently lacked a cell wall lytic enzyme was also fully transformable. A mutant strain that was four times more sensitive to ultraviolet light than the wild type also transformed normally. Recipient cells of this strain were deficient in the repair of ultraviolet-irradiated transforming DNA. Mutants were found which, unlike the wild type, integrated donor markers only with high efficiency, thereby indicating that a particular cellular component that is susceptible to loss by mutation, such as an enzyme, is responsible for low integration efficiency. Images
TL;DR: Three types of mutants were selected in yeast and the mechanisms of their resistance were investigated, finding one is dominant and weakly resistant to 5- fluorouracil, 5-fluorocytosine, and 5- Fluorouridine, and two are recessive and unable to catalyze one of the steps involved in uracil transformation into uridine 5'-monophosphate.
Abstract: Mutants resistant to 5-fluorouracil, 5-fluorocytosine, and 5-fluorouridine were selected in yeast, and the mechanisms of their resistance were investigated. The investigated mutations map in seven different loci. (i) A mutation at the locus FUI 1 gives specifically resistance to 5-fluorouridine. (ii) Two loci are involved in a specific 5-fluorocytosine resistance: a mutation at locus FCY 1 produces a loss of cytosine deaminase activity; a mutation at locus FCY 2 results in the loss of the activity of a cytosine-specific permease. (iii) A mutation at the locus FUR 4 gives a simultaneous resistance to 5-fluorouracil and to 5-fluorouridine by loss in the activity of the uracil-specific permease. (iv) We found three types of mutants in the locus FUR 1. One is dominant and weakly resistant to 5-fluorouracil, 5-fluorocytosine, and 5-fluorouridine. The two others are recessive and are unable to catalyze one of the steps involved in uracil transformation into uridine 5′-monophosphate; this block-age explains their strong resistance to 5-fluorouracil and 5-fluorocytosine. Of these two mutants, one is resistant to 5-fluorouridine and the other is not. (v) Mutations at locus FUR 2 give resistance to 5-fluorouracil, 5-fluorocytosine, and 5-fluorouridine. These mutations are dominant and lead to a loss in the feedback regulation of the aspartic transcarbamylase activity by uridine triphosphate. (vi) The mutants FUR 3 are resistant to 5-fluorocytosine and 5-fluorouridine. They are dominant and physiologically related to the mutants of the locus FUR 1 but their mechanism of resistance is not understood.
TL;DR: The arrangement of species based on ribosomal RNA homologies agrees in most cases with current taxonomic groupings.
Abstract: Evolutionary divergence among species of the yeast genus Saccharomyces was estimated from measurements of deoxyribonucleic acid (DNA)/DNA and ribosomal ribonucleic acid (RNA)/DNA homology. Much diversity was found in the DNA base sequences with several species showing little or no homology to the three reference species, S. cerevisiae, S. lactis, and S. fragilis. These three reference species also showed little or no homology to each other. On the other hand the diversity among ribosomal RNA base sequences was small since most species showed a high degree of homology to the reference species. The arrangement of species based on ribosomal RNA homologies agrees in most cases with current taxonomic groupings. A yeast hybrid (S. fragilis × S. lactis) was shown to contain two nonhomologous genomes. A minimum genome size of 9.2 × 109 daltons for S. cerevisiae was calculated from the rate of DNA renaturation.
TL;DR: Partially purified nitrofurazone reductase preparations catalyze the conversion of nitro furazone to compounds which bind to protein and are not removed by prolonged dialysis against 8 m urea or by cold acid.
Abstract: When Escherichia coli strain B/r is exposed to 10 to 20 μg of nitrofurazone per ml, mutants with roughly threefold resistance are obtained. Treatment of these mutants with higher concentrations of nitrofurazone yields strains with six- to seven-fold resistance over strain B/r. Each of these steps toward nitrofurazone resistance is accompanied by loss of soluble nitrofurazone reductase activity. When sensitive bacteria are exposed to labeled nitrofurazone or labeled 2-nitrofuran, a considerable amount of radioactivity becomes bound to the cold trichloroacetic acid-insoluble fraction. Very little activity becomes bound in the mutants with six- to seven-fold resistance; mutants with intermediate resistance show intermediate levels of binding. Partially purified nitrofurazone reductase preparations catalyze the conversion of nitrofurazone to compounds which bind to protein and are not removed by prolonged dialysis against 8 m urea or by cold acid. Nitrofurazone reduced by xanthine oxidase or electrolytically reduced also yields compounds which react with protein to form stable derivatives.
TL;DR: Transduction experiments showed that the genes for enterobactin biosynthesis are closely linked on the S. typhimurium chromosome, and it is suggested that they form an operon which is repressed by the presence of iron.
Abstract: A number of mutants of Salmonella typhimurium were isolated which are blocked in the biosynthesis of enterobactin, an iron chelator that is secreted by the wild-type bacteria when they are grown on low iron media One class of these enb mutants accumulates the enterobactin precursor 2,3-dihydroxybenzoic acid, and another class does not accumulate any detectable catechol precursor The enb mutants grow very well on a glucose-mineral salts medium Addition of citrate, itself an iron chelator, to the medium drastically inhibits growth unless the medium is supplemented with enterobactin or iron salts Citrate inhibits iron uptake from the medium by enb mutants; enterobactin counteracts this inhibition and also, by itself, increases iron uptake Thus, the apparent function of enterobactin is to promote the absorption of iron from the medium by the bacteria Transduction experiments showed that the genes for enterobactin biosynthesis are closely linked on the S typhimurium chromosome It is suggested that they form an operon which is repressed by the presence of iron S typhimurium can utilize the iron chelate ferrichrome (Deferriferrichrome is a cyclic hexapeptide that is produced by some fungi but not by S typhimurium) The enb mutants use ferrichrome as an effective growth factor
TL;DR: The ultrastructure of Mycoplasma pneumoniae cultivated in broth on glass and plastic surfaces was studied by scanning and transmission electron microscopy and sectioned organisms were seen to contain ribosome-like structures.
Abstract: The ultrastructure of Mycoplasma pneumoniae cultivated in broth on glass and plastic surfaces was studied by scanning and transmission electron microscopy. The organisms grew as filaments, which by over-crossing eventually formed a dense network on the surface and in colonies composed mainly of rounded and elongated forms. The filaments were usually thinner at the ends and terminated with a knob-like structure. Some filaments possessed short ramifications which also ended with a knob, and others showed constrictions. Sectioned organisms were seen to contain ribosome-like structures. Many organisms had a specialized structure at their thinner end, which consisted of a dense rod surrounded by electron-lucent cytoplasm and ending with a platelike thickening.
TL;DR: In this paper, the cells of least or greatest density after banding in Renografin-sucrose density gradients are selected from asynchronously growing cell cultures.
Abstract: Yeast cells undergo periodic fluctuations in density during the cell division cycle such that a minimum in density occurs at the time of cell separation whereas a maximum occurs between the time of deoxyribonucleic acid replication and nuclear division. Synchronous cultures can be selected from asynchronously growing cell cultures by withdrawing the cells of least or greatest density after banding in Renografin-sucrose density gradients. This technique is rapid, reproducible, and almost unlimited in capacity.
TL;DR: An envelope preparation containing the cell wall and cytoplasmic membrane of Escherichia coli was obtained by breaking the cells with a French pressure cell and sedimentating the envelope fraction by ultracentrifugation, suggesting that this organism is able to adapt to changes in growth environment with only minor modifications of the major proteins of the cell envelope.
Abstract: An envelope preparation containing the cell wall and cytoplasmic membrane of Escherichia coli was obtained by breaking the cells with a French pressure cell and sedimentating the envelope fraction by ultracentrifugation. This fraction was prepared for polyacrylamide gel electrophoresis by dissolving the protein in an acidified N,N′-dimethylformamide, removing lipids by gel filtration in the same organic solvent and removing the solvent by dialysis against aqueous urea solutions. More than 80% of the total protein of the envelope fraction was recovered in soluble form. Electrophoresis on sodium dodecyl sulfate-containing gels yielded from 20 to 30 well-resolved bands of protein. One major protein band was observed on the gels. This protein had a molecular weight of 44,000 and accounted for as much as 40% of the total protein of the envelope fraction. A double-labeling technique was used to examine the protein composition of the envelope fraction from cells grown under different sets of conditions which result in large changes in the levels of membrane-bound oxidative enzymes. These changes in growth conditions resulted in only minor alterations in the protein profiles observed on the gels, suggesting that this organism is able to adapt to changes in growth environment with only minor modifications of the major proteins of the cell envelope.
TL;DR: The antibiotic polyoxin D was shown to inhibit the incorporation of (14)C-glucosamine into cell wall chitin in Neurospora crassa at levels which were comparable with those required for inhibition of fungal growth.
Abstract: The antibiotic polyoxin D was shown to inhibit the incorporation of (14)C-glucosamine into cell wall chitin in Neurospora crassa at levels which were comparable with those required for inhibition of fungal growth. At the same time, the antibiotic increased the accumulation of a nucleotide, which was identified as uridine diphosphate (UDP)-N-acetylglucosamine, indicating inhibition of chitin synthesis. Chitin synthetase (UDP-N-acetylglucosamine: chitin N-acetylglucosaminyl transferase, EC 2.4.1.16) of N. crassa was found to be strongly inhibited by polyoxin D, as determined by the transfer of (14)C-N-acetylglucosamine from (14)C-UDP-N-acetylglucosamine to the particulate fraction. The inhibition was competitive with respect to UDP-N-acetylglucosamine and specific for chitin synthetase. The K(i) for polyoxin D in the reaction was 1.40 x 10(-6)m, and the K(m) for UDP-N-acetylglucosamine was 1.43 x 10(-3)m. The formation of osmotically sensitive, protoplast-like structures, when the fungus Cochliobolus miyabeanus was grown in the presence of polyoxin D, also suggested that the primary site of action of polyoxin D was in the formation of cell wall structures.
TL;DR: Although the phosphoenolpyruvate phosphotransferase system is an important system for the transport of sugars in bacteria carrying out anaerobic glycolysis, it plays no role in sugar transport by those organisms having a strictly oxidative physiology.
Abstract: A survey of the occurrence of the phosphoenolpyruvate-dependent glucose phosphotransferase system was carried out in a number of bacteria, representing both gram-positive and gram-negative facultative anaerobic and strictly aerobic types The system was found to be present in representatives of genera that are characteristically facultative anaerobes, but the system was absent in members of those genera that are strictly aerobic Thus, although the phosphoenolpyruvate phosphotransferase system is an important system for the transport of sugars in bacteria carrying out anaerobic glycolysis, it plays no role in sugar transport by those organisms having a strictly oxidative physiology A fundamentally different system, probably not involving phosphorylation during transport, is indicated in this latter group
TL;DR: Continuous observation of the lytic process under a phase-contrast microscope suggested that a close contact between the polar tip of the myxobacter and the alga is necessary for lysis.
Abstract: Enrichment from local fishponds led to the isolation of a bacterium capable of lysing many species of unicellular and filamentous blue-green algae, as well as certain bacteria. The isolate is an aflagellate, motile rod which moves in a gliding, flexuous manner; the organism is capable of digesting starch and agar, but not cellulose and gelatin. Its deoxyribonucleic acid base pair composition (per cent guanine plus cytosine approximately 70) shows a close resemblance to that of the fruiting myxobacteria. Algae in lawns on agar plates were lysed rapidly by the myxobacter, but only limited and slow lysis occurred in liquid media, and no lysis took place when liquid cultures were shaken. No diffusible lytic factors would be demonstrated. Continuous observation of the lytic process under a phase-contrast microscope suggested that a close contact between the polar tip of the myxobacter and the alga is necessary for lysis. The lytic action is limited to the vegetative cells of the algae, whereas heterocysts are not affected. The gas vacuoles of the algal host are the only remnant visible after completion of digestion by the myxobacter.
TL;DR: In exponentially growing and dividing cells of Streptococcus faecalis, it is proposed that the leading edge of the annularly closing cross wall is the point of extension for both cross wall and peripheral wall.
Abstract: In exponentially growing and dividing cells of Streptococcus faecalis, it is proposed that the leading edge of the annularly closing cross wall is the point of extension for both cross wall and peripheral wall. Peripheral wall extension is thought to be produced by the separation or splitting of the cross wall at its junction with peripheral wall. This results in the pushing of the equatorial wall bands, found on S. faecalis walls, to subsequatorial positions. These bands therefore mark the separation of old wall from new wall. Mesosomal formation was observed usually to precede cross wall initiation.
TL;DR: The desensitization of glycerol kinase to feedback inhibition enhances the power of Glycerol to exert catabolite repression, both on the enzymes of the glycerl system itself and on those of the lactose system, but does not eliminate the phenomenon of diauxic growth in a glucose-glycerol medium.
Abstract: The activity of glycerol kinase is rate-limiting in the metabolism of glycerol by cells of Escherichia coli. A mutant strain producing a glycerol kinase resistant to inhibition by fructose-1,6-diphosphate grows faster than its wild-type parent on glycerol as the sole source of carbon and energy. The amount of intracellular fructose-1,6-diphosphate was determined for wild-type cells growing exponentially on glycerol. The water content of such cells was also determined, allowing calculation of the intracellular concentration of fructose-1,6-diphosphate. This value, 1.7 mm, is adequate to exert substantial inhibition on the wild-type glycerol kinase. The desensitization of glycerol kinase to feedback inhibition also enhances the power of glycerol to exert catabolite repression, both on the enzymes of the glycerol system itself and on those of the lactose system. However, desensitization of glycerol kinase alone does not eliminate the phenomenon of diauxic growth in a glucose-glycerol medium. Biphasic growth in such a medium is abolished if the altered enzyme is produced constitutively. The constitutive production of the mutant kinase at high levels, however, renders the cells vulnerable to glycerol. Thus, when the cells have been grown on a carbon source with a low power for catabolite repression, e.g., succinate, sudden exposure to glycerol leads to overconsumption of the nutrient and cell death.
TL;DR: Experimental support is provided for the view that the large majority of the established Mycoplasma species require cholesterol for growth.
Abstract: Cholesterol requirement for growth of mycoplasmas was tested in a serum-free medium supplemented with albumin, l-arginine, palmitic acid, and various concentrations of cholesterol dissolved in Tween 80. In cases in which Tween 80 was shown to inhibit growth, the test medium was supplemented with cholesterol dissolved in ethanol. Of the 31 species examined, all but Mycoplasma laidlawii, M. granularum, and Mycoplasma species strain S-743 exhibited a growth response to cholesterol. No requirement for cholesterol could be shown with the stable L-phase variants of Streptobacillus moniliformis and Proteus species. The results provide experimental support for the view that the large majority of the established Mycoplasma species require cholesterol for growth.
TL;DR: A thermophilic (70 C), gram-negative bacterium has been isolated that resembles Thermus aquaticus but lacks the type-characteristic yellow carotenoid.
Abstract: A thermophilic (70 C), gram-negative bacterium has been isolated that resembles Thermus aquaticus but lacks the type-characteristic yellow carotenoid.