bluuebunny/crossref_metadata_2025 huggingface dataset:
Role and control of isocitrate lyase in Candida lipolytica
Authors: Matsuoka M , Ueda Y , Aiba S
Published: January 2020

Mutants of Candida lipolytica that were unable to grow on acetate but able to utilize succinate or glycerol as a sole carbon source were isolated. Amongst the mutants isolated, one strain (Icl-) was specifically deficient in isocitrate lyase activity, whereas another strain (Acos-) was deficient in acetyl coenzyme A synthetase activity. Since the Icl- mutant could not grow either on n-alkane or its derivatives, such as fatty acid and long-chain dicarboxylic acid, any anaplerotic route other than the glyoxylate pathway was inconceivable as far as growth on these carbon sources was concerned. Acetyl coenzyme A is most likely a metabolic inducer of isocitrate lyase and malate synthase, because the Acos- mutant was characterized by the least susceptibility to induction of these enzymes by acetate. The structural gene for isocitrate lyase was most probably impaired in the Icl- mutant, since revertants (Icl-) produced thermolabile isocitrate lyase. The production of isocitrate from n-alkane by the revertants was enhanced in comparison with the parental strain.
Yeast-phase cell cycle of the polymorphic fungus Wangiella dermatitidis
Authors: Roberts R L , Szaniszlo P J
Published: January 2020

The yeast-phase cell cycle of Wangiella dermatitidis was studied using flow microfluorimetry and the deoxyribonucleic acid (DNA) synthesis inhibitor hydroxyurea (HU). Exposure of exponential-phase yeastlike cells to 0.1 M HU for 3 to 6 h resulted in the arrest of the cells in DNA synthesis and produced a nearly homogeneous population of unbudded cells. Treatment of the yeast-phase cells with HU for 9 h or longer resulted in the accumulation of the cells predominantly as budded forms having either a single nucleus in the mother cell or a single nucleus arrested in the isthmus between the mother cell and the daughter bud. Exposure of unbudded stationary-phase cells to 0.1 M HU resulted in the accumulation of the cells in the same phenotypes. Analysis by flow microfluorimetry and cell counts of HU-inhibited mithramycin-stained cells indicated that the eventual progress of HU-inhibited cells from unbudded to the two budded forms was due to the limited continuation of the growth sequence of the cell cycle even in the absence of DNA synthesis, nuclear division, and in some cases nuclear migration. On the basis of these observations and the results of flow microfluorimetric analysis of exponential-phase cells, a map of the yeast-phase cell cycle was constructed. The cycle appears to consist of two independent sequences of events, a budding growth sequence and a DNA division sequence. The nuclear division cycle of yeast-phase cells growing exponentially with a 4.5-h generation time is composed of a G1 interval of 148 min, as S phase of 16 min, and a G2 plus M interval of 107 min.
Tetracyclin-stimulated expression of ampicillin resistance in Haemophilus influenzae
Authors: Stuy J H
Published: January 2020

Tetracycline at a low concentration stimulated the expression of ampicillin resistance in certain strains of Haemophilus influenzae.
Postreplication repair of deoxyribonucleic acid and daughter strand exchange in uvr- mutants of Bacillus subtilis
Authors: Dodson L A , Hadden C T
Published: January 2020

The fate of pyrimidine dimers in deoxyribonucleic acid (DNA) newly synthesized by Bacillus subtilis after ultraviolet irradiation was monitored by use of a damage-specific endonuclease that introduces single-strand breaks adjacent to nearly all of the dimer sites. Two Uvr- strains, one defective in the initiation of dimer excision and the other defective in a function required for efficient dimer excision, were found to be similar to their wild-type parent in the kinetics and extent of converting low-molecular-weight DNA newly synthesized after ultraviolet irradiation to high molecular weight. In the Uvr- strains large molecules of newly synthesized DNA remained susceptible to nicking by the damage-specific endonuclease even after extended incubation in growth medium, whereas the enzyme-sensitive sites were rapidly removed from both preexisting and newly synthesized DNA in Uvr+ cells. Our results support the hypothesis that postreplication repair in bacteria includes recombination between dimer-containing parental DNA strands and newly synthesized strands.
Nutritional factors controlling exocellular protease production by Pseudomonas aeruginosa
Authors: Jensen S E , Fecycz I T , Campbell J N
Published: January 2020

A defined medium capable of supporting growth and exocellular protease production by clinical isolates of Pseudomonas aeruginosa has been developed. Control of protease production is effected by a mixture of three amino acids and glucose.
Alteration of Escherichia coli murein during amino acid starvation
Authors: Goodell W , Tomasz A
Published: January 2020

We have studied the mechanisms by which amino acid starvation of Escherichia coli induces resistance against the lytic and bactericidal effects of penicillin. Starvation of E. coli strain W7 of the amino acids lysine or methionine resulted in the rapid development of resistance to autolytic cell wall degradation, which may be effectively triggered in growing bacteria by a number of chemical or physical treatments. The mechanism of this effect in the amino acid-starved cells involved the production of a murein relatively resistant to the hydrolytic action of crude murein hydrolase extracts prepared from normally growing E. coli. Resistance to the autolysins was not due to the covalently linked lipoprotein. Resistance to murein hydrolase developed most rapidly and most extensively in the portion of cell wall synthesized after the onset of amino acid starvation. Lysozymes digests of the autolysin-resistant murein synthesized during the first 10 min of lysine starvation yielded (in addition to the characteristic degradation products) a high-molecular-weight material that was absent from the lysozyme-digests of control cell wall preparations. It is proposed that inhibition of protein synthesis causes a rapid modification of murein structure at the cell wall growth zone in such a manner that attachment of murein hydrolase molecules is inhibited. The mechanism may involve some aspects of the relaxed control system since protection against penicillin-induced lysis developed much slower in amino acid-starved relaxed controlled (relA) cells than in isogenic stringently controlled (relA+) bacteria.
Surface projections of a chlamydia-like parasite of midge larvae
Authors: Morel G
Published: January 2020

Projections are described in a chlamydia-like organism similar to Rickettsiella chironomi. These projections were present only on the edge of the disk-like elementary bodies; rod-shaped, they were found to be composed of repetitive subunits.
Nature, type of linkage, quantity, and absolute configuration of (3-hydroxy) fatty acids in lipopolysaccharides from Bacteroides fragilis NCTC 9343 and related strains
Authors: Wollenweber H W , Rietschel E T , Hofstad T , Weintraub A , Lindberg A A
Published: January 2020

The main fatty acids present in lipopolysaccharides from Bacteroides fragilis NCTC 9343 were identified as 13-methyl-tetradecanoic, D-3-hydroxypentadecanoic, D-3-hydroxyhexadecanoic, D-3-hydroxy-15-methyl-hexadecanoic, and D-3-hydroxyheptadecanoic acids. Of these, 13-methyl-tetradecanoic acid is exclusively ester bound, and 3-hydroxy-15-methyl-hexadecanoic acid is exclusively involved in amide linkage. The other 3-hydroxy fatty acids are both ester and amide bound. All 3-hydroxy fatty acids possess the D configuration, and the 3-hydroxyl group of ester-linked 3-hydroxy fatty acids is not substituted. Lipopolysaccharides of related Bacteroides species (B. thetaiotaomicron, B. ovatus, B. distasonis, and B. vulgatus) showed a fatty acid spectrum with both similar and distinct features compared to that of B. fragilis lipopolysaccharides.
Location of two relaxation nick sites in R6K and single sites in pSC101 and RSF1010 close to origins of vegetative replication: implication for conjugal transfer of plasmid deoxyribonucleic acid
Authors: Nordheim A , Hashimoto-Gotoh T , Timmis K N
Published: January 2020

A nick-labeling method has been used to localize the relaxation complex nick sites in three plasmids (pSC101, RSF1010, and R6K) that differ markedly in their host range, deoxyribonucleic acid replication, and conjugal transfer properties. Single specific relaxation sites were located in pSC101 and RSF1010, but surprisingly two distinct sites could be identified in the bi-origin plasmid R6K. In all cases, relaxation nick sites, which are thought to be origins of plasmid conjugal transfer, were shown to be located near origins of vegetative replication. This result suggests a functional interaction between these two types of deoxyribonucleic acid loci, and we speculate here that application events initiated at origins of replication may constitute an integral part of the process of conjugal transfer of small plasmids among bacteria. Consistent with this proposal is the finding that inhibition of vegetative replication of the pSC101 and ColE1 plasmids results in a severe inhibition of their conjugal transfer ability.
Effect of glucose on alpha-glucan degradation in Polyporus circinatus
Authors: Oliveira M B , Zancan G T
Published: January 2020

The effect of glucose on the enzymes involved in the degradation of a reserve alpha-glucan in Polyporus circinatus was studied. The levels of phosphorylase activity, endoamylase, amylo-1,6-glucosidase were regulated by glucose concentration.
Deletion map of the Escherichia coli structural gene for alkaline phosphatase, phoA
Authors: Sarthy A , Michaelis S , Beckwith J
Published: January 2020

Lambda transducing phages containing portions of the phoA gene have been isolated and used to construct a deletion map of the phoA gene. The isolation of a plaque-forming lambda transducing phage carrying the entire phoA gene is also described. Two new methods for screening or selection of mutants that have altered levels of alkaline phosphatase activity are reported.
Use of gene fusions to determine the orientation of gene phoA on the Escherichia coli chromosome
Authors: Sarthy A , Michaelis S , Beckwith J
Published: January 2020

We present genetic evidence which demonstrates that the phoA gene is transcribed in the clockwise direction on the Escherichia coli chromosome, in contrast to an earlier proposal. Our conclusion is based on analysis of various genetic fusions between the lac operon and the phoA gene.
Genetic variability in Halobacterium halobium
Authors: Pfeifer F , Weidinger G , Goebel W
Published: January 2020

Halobacterium halobium exhibits an extraordinary degree of spontaneous variability. Mutants which are defective in the formation of gas vacuoles (vac) arise at a frequency of 10(-2). Other easily detectable phenotypes, like the synthesis of bacterioruberin (Rub) or the synthesis of retinal (Ret) and bacterio-opsin (Ops), the two components which form the purple membrane (Pum) of H. halobium, are lost at a frequency of about 10(-4). With the same frequency a mutant type appears which exhibits an extremely high variability in these phenotypes. With the exception of the ret mutants, all spontaneously arising mutants show alterations, i.e., insertions, rearrangements, or deletions, in the plasmid pHH1. It appears that the introduction of one insertion into pHH1 triggers further insertions, which makes the identification of relationships between phenotypic and genotypic alterations rather difficult. From the analysis of a large number of spontaneous vac mutants and their vac+ revertants it can be concluded that the formation of the gas vacuoles is determined or controlled by plasmid genes. No such conclusion is yet possible for the rub mutants, although all mutants of this type so far analyzed exhibit a defined insertion. pum mutants which have lost the capability of forming bacterio-opsin carry insertions in the plasmid which are distributed over a rather large region of the plasmid. No strains of H. halobium could be obtained which had lost plasmid pHH1 completely.
Induction of Bacillus subtilis sporulation by nucleosides: inosine appears to be sporogen
Authors: Sekar V , Wilson S P , Hageman J H
Published: January 2020

Inosine completely reversed the selective inhibition of sporulation in Bacillus subtilis 168 caused bym-aminobenzeneboronic acid; guanosine and adenosine, but not xanthosine, partially reversed inhibition, whereas pyrimidine nucleosides were slightly effective. In addition, 0.005 to 0.025 mM inosine caused a four- to fivefold stimulation of sporulation of B. subtilis grown in minimal salts medium. Ultraviolet and infrared spectra and other physical and chemical properties of inosine were markedly similar to those of "sporogen," a previously described endogenous sporogenesis factor present in sporulating Bacillus species.
Genetics of ribosomal components in Podospora spp. as revealed by interspecific hybridization
Authors: Perrot M , Begueret J
Published: January 2020

The ribosomal proteins of several species within the genus Podospora have been analyzed by two-dimensional polyacrylamide gel electrophoresis. Differences in the migration of four proteins of the large subunit have been found between the two hybridizable species Podospora anserina and P. comata. Haploid offspring of this interspecific cross show all possible combinations of ribosomal proteins from both species. Therefore, the homologous proteins are functionally interchangeable. Moreover, the sizes of the different classes of offspring show that the genes encoding the four ribosomal proteins L2, L13, L16, and L20 are not clustered.
Regulatory mutations conferring constitutive synthesis of major outer membrane proteins (OmpC and OmpF) in Escherichia coli
Authors: Sato T , Yura T
Published: January 2020

An ompB strain of Escherichia coli K-12 lacking major outer membrane proteins OmpC and OmpF was used to isolate a pair of mutants that have restored the ability to synthesize either OmpC or OmpF protein. These mutants were found to produce the respective proteins constitutively under the several conditions where the synthesis in the wild-type strain was markedly repressed; namely, in the absence of the ompB gene function, under restrictive medium conditions, or upon lysogenization with phage PA-2. The mutations ompCp1 and ompFp9 responsible for such synthesis were shown to be located in the close vicinity of the corresponding structural genes, ompC and ompF. Moreover, the mutations affect the expression of these genes in a cis-dominant fashion. Taken together with other evidence, it was suggested that ompCp1 and ompFp9 represent regulatory site mutations occurring at the promoter regions of ompC and ompF respectively. Relevance of these findings to the genetic control of outer membrane protein synthesis is discussed.
Characterization and mapping of temperature-sensitive division initiation mutations of Bacillus subtilis
Authors: Callister H , Wake R G
Published: January 2020

Two temperature-sensitive, filamenting mutants of Bacillus subtilis (ts1 and ts12) have been shown to be defective in the initiation of septation. Recombination index mapping showed that these mutations mapped in two different but closely linked genes. A third proposed initiation mutation, tms-12, probably maps in the same gene as ts12. Another proposed initiation mutation was not linked with these genes by transformation, indicating that there was a minimum of three genes involved in the initiation of division. PBS1 transduction mapping located these three genes close to the pyr cluster.
sfrA and sfrB products of Escherichia coli K-12 are transcriptional control factors
Authors: Beutin L , Manning P A , Achtman M , Willetts N
Published: January 2020

The mechanisms whereby mutations in Escherichia coli K-12 genes sfrA and sfrB reduce expression of the transfer functions of sex factor F have been examined by assaying the levels of tra messenger ribonucleic acid and of tra proteins. The sfrA product was necessary for efficient transcription of the control gene traJ and, directly or indirectly, for transcription of the traY leads to Z operon. In the absence of sfrA, reduced levels of the traJ and traT proteins were observed in the outer membrane. The sfrB product was needed to prevent premature transcription at one or more rho-dependent termination sites. sfrB mutations also reduced synthesis of full-length lipopolysaccharide molecules, of several chromosomally determined outer membrane proteins, and of functional flagella. Thus, the sfrB product may act as an antiterminator in transcription of several operons encording cell envelope components.
Escherichia coli heat-labile enterotoxin genes are flanked by repeated deoxyribonucleic acid sequences
Authors: Yamamoto T , Yokota T
Published: January 2020

The enterotoxin regions of the heat-labile and heat-stable enterotoxin (LT+ ST+) plasmid, pJY11, originating in a clinically isolated Escherichia coli strain, have been isolated as various-sized deoxyribonucleic acid (DNA) fragments by using cloning vehicles. The structure of the LT+ region and its neighboring DNA regions was studied by utilizing these recombinant plasmids. The LT+ region consisted of at least two genes, toxA and toxB, which could complement each other in trans. The toxA- and toxB-encoded polypeptides (LT subunits A and B, respectively) were identified by their immunological cross-reactivity with Vibrio cholerae enterotoxin subunit A or B. These tox genes and the promoter(s) were localized with respect to the restriction endonuclease cleavage map. The LT+ region was flanked by repeated DNA sequences (designated as beta). Another tox gen(s), encoding ST (designated as toxS), which was also flanked by inverted, repeated DNA sequences (designated as alpha), was located between one of the beta sequences and the LT+ region. These novel DNA structures (beta-alpha-toxS-alpha-toxA-toxB-beta) suggest the possibility that the LT+ region is on a transposon containing an ST transposon within the structure.
Comparative studies of two membrane fractions isolated from chemotrophically and phototrophically grown cells of Rhodopseudomonas capsulata
Authors: Garcia A F , Drews G , Reidl H H
Published: January 2020

Light and heavy membrane fractions have been isolated by equilibrium sucrose density centrifugation from Rhodopseudomonas capsulata 938 GCM grown aerobically in the dark (chemotrophically) and anaerobically in the light (phototrophically). The densities of the light and heavy fractions from phototrophic cells were 1.1004 to 1.1006 and 1.1478, respectively, and the densities of the light and heavy fractions from chemotrophic cells were 1.0957 to 1.0958 and 1.1315, respectively. Both fractions were active in photochemical and respiratory functions and in electron transport-coupled phosphorylation. The light membrane fraction isolated from chemotrophic cells contained the reaction center and the light-harvesting pigment-protein complex B 870, but not the variable light-harvesting complex B 800-850. A small amount of the complex B 800-850 was present in the light fraction isolated from phototrophically grown cells, but it was not energetically coupled to the photosynthetic apparatus. From inhibitor studies, difference spectroscopy, and measurement of enzyme activities it was tentatively concluded that the light membrane fraction contains only the reduced nicotinamide adenine dinucleotide-oxidizing electron transport chain having a KCN-insensitive, low-potential cytochrome c oxidase, whereas the heavy fraction contains additionally the succinate dehydrogenase and a high-potential cytochrome b terminal oxidase sensitive to KCN. The light membrane fraction was more labile than the heavy fraction in terms of phosphorylating activity.
Rifampin resistance mutations that alter the efficiency of transcription termination at the tryptophan operon attenuator
Authors: Yanofsky C , Horn V
Published: January 2020

Rifampin-resistant mutants of Escherichia coli were isolated which had altered patterns of resistance or sensitivity to the inhibitory compounds 5-methyltryptophan and 5-methylanthranilate. The levels of tryptophan (trp) operon polypeptides in different rifampin-resistant mutants were elevated or reduced, in a manner consistent with their sensitivity to the two analogs. Complementation tests established that the mutations were in rpoB, the structural gene for the beta subunit of ribonucleic acid polymerase. Introduction of these rpoB mutations into mutant strains which terminate transcription abnormally at the trp operon attenuator established that the rpoB mutations alter trp operon expression by increasing or decreasing transcription termination at the attenuator. The rpoB mutations affected transcription termination at the attenuator only in strains which were able to form what is thought to be a ribonucleic acid termination structure. These findings suggest that alteration of the beta subunit of ribonucleic acid polymerase directly or indirectly affects ribonucleic acid polymerase's recognition of the transcription termination signal at the trp operon attenuator.
Inactivation of the ribonucleic acid-processing enzyme ribonuclease E blocks cell division
Authors: Goldblum K , Apririon D
Published: January 2020

The Escherichia coli endoribonuclease ribonuclease E, the enzyme responsible for the processing of precursor 5S ribosomal ribonucleic acid (RNA) from the nascent ribosomal RNA transcript, is thermolabile in rne-3071 mutants. When cells of such a strain were shifted to a nonpermissive temperature, the function of ribonuclease E was almost instantaneously inactivated. However, a threefold linear increase in absorbance took place over a 3-h period, and similar linear increases occurred in all the macromolecules we measured, including deoxyribonucleic acid, RNA, protein, and lipopolysaccharides. Interestingly, during this period, the cells elongated but failed to divide. Thus, these experiments suggest that an early effect of inactivation of the RNA processing enzyme ribonuclease E is a block in cell division.
Ethanol production by thermophilic bacteria: metabolic control of end product formation in Thermoanaerobium brockii
Authors: Ben-Bassat A , Lamed R , Zeikus J G
Published: January 2020

Specific changes in the chemical and microbial composition of Thermoanaerobium brockii fermentations were compared and related to alterations of process rates, end product yields, and growth parameters. Fermentation of starch as compared with glucose was associated with significant decreases in growth rate and intracellular fructose-1,6-bisphosphate concentration and with a dramatic increase in the ethanol/lactate product ratio. Glucose or pyruvate fermentation in the presence of acetone was correlated with increased substrate consumption, growth (both rate and yield), acetate yield, and quantitative reduction of acetone to isopropanol in lieu of normal reduced fermentation products (i.e., H2, ethanol, lactate). Acetone altered pyruvate phosphoroclastic activity of cell extracts in that H2, lactate, and ethanol levels decreased, whereas the acetate concentration increased. Glucose fermentation in the presence of exogenous hydrogen was associated with inhibition of endogenous H2 production and either increased ethanol/acetate product ratios and decreased growth at less than 0.5 atm (51 kPa) of H2 or total growth inhibition at 1.0 atm (102 kPA). The effects of exogenous hydrogen on glucose fermentation were totally reversed by the addition of acetone. Glucose fermentation in coculture with Methanobacterium thermoautotrophicum correlated with increased growth (both rate and yield), acetate yield, and the formation of methane in lieu of monoculture reduced products. In coculture, but not monoculture, T. brockii grew on ethanol as the energy source, and acetate and methane were the end products as a direct consequence of hydrogen consumption by the methanogen.
Ribonucleic acid synthesis during fruiting body formation in Myxococcus xanthus
Authors: Smith B A , Dworkin M
Published: January 2020

A method has been devised that allowed us, for the first time, to pulse-label M. xanthus cells with precursors for ribonucleic acid biosynthesis while they were undergoing fruiting body formation. Using this method, we examined patterns of ribonucleic acid (RNA) accumulation throughout the process of fruiting body formation. As development proceeded, the rate of RNA accumulation increased at two periods of the developmental cycle: once just before aggregation and once late in the cycle, when sporulation was essentially completed. In contrast to vegetatively growing cells, in which only stable RNA species are labeled during a 30-min pulse, the majority of radioactivity found in RNA from 30-min pulse-labeled developing cells was found in an unstable heterodisperse fraction that migrated to the 5S to 16S region of sucrose density gradients and sodium dodecyl sulfate-polyacrylamide gels. This pattern of incorporation could not be induced (i) by a shift down of vegetatively growing cells to a nutritionally poor medium, in which the generation time was increased to that of developing cells during the growth phase, or (ii) by plating of vegetative cells onto the same solid-surface environment as that of developing cells, but which surface supported vegetative growth rather than fruiting body formation. Thus, the RNA synthesis pattern observed appeared to be related to development per se rather than to nutritional depletion or growth on a solid surface alone. The radioactivity incorporated into the unstable 5S to 16S RNA fraction accumulated as the pulse length was increased from 10 to 30 min; in contrast, an analogous unstable fraction from vegetative cells decreased as pulse length was increased. This suggested that developmental 5S to 16S RNA was more stable than vegetative cell 5S to 16S RNA (presumptive messenger RNA). However, during a 45-min chase period, radioactivity in 30-min-pulse-labeled developmental 5S to 16S RNA decayed to an extent twice that of developmental RNA located in 16S and 23S regions of sucrose density gradients and was considerably less stable than the 5S, 16S, and 23S RNA species labeled during a 30-min pulse of vegetative cells.
Effects of aerobiosis and nitrogen source on the proton motive force in growing Escherichia coli and Klebsiella pneumoniae cells
Authors: Kashket E R
Published: January 2020

The electrochemical gradient of hydrogen ions, or proton motive force (PMF), was measured in growing Escherichia coli and Klebsiella pneumoniae in batch culture. The electrical component of the PMF (delta psi) and the chemical component (delta pH) were calculated from the cellular accumulation of radiolabeled tetraphenylphosphonium, thiocyanate, and benzoate ions. In both species, the PMF was constant during exponential phase and decreased as the cells entered stationary phase. Altering the growth rate with different energy substrates had no effect on the PMF. The delta pH (alkaline inside) varied with the pH of the culture medium, resulting in a constant internal pH. During aerobic growth in media at pH 6 to 7, the delta psi was constant at 160 mV (negative inside). The PMF, therefore, was 255 mV in cells growing at pH 6.3, and decreased progressively to 210 mV in pH 7.1 cultures. K. pneumoniae cells and two E. coli strains (K-12 and ML), including a mutant deficient in the H+-translocating ATPase and a pleiotropically energy-uncoupled mutant with a normal ATPase, had the same PMF during aerobic exponential phase. During anaerobic growth, however, both species had delta psi values equal to 0. Therefore, the PMF in anaerobic cells consisted only of the delta pH component, which was 75 mV or less in cells growing at pH 6.2 or greater. These data thus met the expectation that cells deriving metabolic energy from respiration have a PMF above a threshold value of about 200 mV when the ATPase functions in the direction of H+ influx and ATP synthesis; in fermenting cells, a PMF below a threshold value was expected since the enzyme functions in the direction of H+ extrusion and ATP hydrolysis. K. pneumoniae cells growing anaerobically had no delta psi whether the N source added was N2, NH+4 or one of several amino acids; the delta pH was unaffected. Therefore, any energy cost incurred by the process of nitrogen fixation could not be detected as an alteration of the proton gradient.
Mutations to tolerance and resistance to pesticin and colicins in Escherichia coli phi
Authors: Ferber D M , Fowler J M , Brubaker R R
Published: January 2020

The universal colicin-indicator strain Escherichia coli phi, unlike E. coli strain K-12, is sensitive to pesticin, a bacteriocin produced by wild-type Yersinia pestis. Eleven distinct phenotypes of E. coli phi mutants were obtained by selection for insensitivity to pesticin, group B colicins, the group A colicin S4, or coliphage T5. Representative isolates from eight of these classes closely resembled resistant receptor mutants (Cir-, Fep-, and TonA-) or tolerant mutants (TonB-, ExbB-, ExbC-, Ivt-, and Cmt-) described in Escherichia coli K-12. The remainder were unique; of these, one resembled TonB- but was also tolerant to colicin S4 (TonB/S4-), and the others exhibited specific resistance to either colicin S4 (Sfr-) or to pesticin (Psr-). All receptor mutants except Psr- remained sensitive to pesticin, whereas TonB/S4, TonB-, ExbB-, and ExbC- isolates were highly tolerant to this bacteriocin.
A family of r-determinants in Streptomyces spp. that specifies inducible resistance to macrolide, lincosamide, and streptogramin type B antibiotics
Authors: Fujisawa Y , Weisblum B
Published: January 2020

Inducible resistance to macrolide, lincosamide, and streptogramin type B antibiotics in Streptomyces spp. comprises a family of diverse phenotypes in which characteristic subsets of the macrolide-lincosamide-streptogramin antibiotics induce resistance mediated by mono- or dimethylation of adenine, or both, in 23S ribosomal ribonucleic acid. In these studies, diverse patterns of induction specificity in Streptomyces and associated ribosomal ribonucleic acid changes are described. In Streptomyces fradiae NRRL 2702 erythromycin induced resistance to vernamycin B, whereas in Streptomyces hygroscopicus IFO 12995, the reverse was found: vernamycin B induced resistance to erythromycin. In a Streptomyces viridochromogenes (NRRL 2860) model system studied in detail, tylosin induced resistance to erythromycin associated with N6-monomethylation of 23S ribosomal ribonucleic acid, whereas in Staphylococcus aureus, erythromycin induced resistance to tylosin mediated by N6-dimethylation of adenine. Inducible macrolide-lincosamide-streptogramin resistance was found in S. fradiae NRRL 2702 and S. hygroscopicus IFO 12995, which synthesize the macrolides tylosin and maridomycin, respectively, as well as in the lincosamide producer Streptomyces lincolnensis NRRL 2936 and the streptogramin type B producer Streptomyces diastaticus NRRL 2560. A wide range of different macrolides including chalcomycin, tylosin, and cirramycin induced resistance when tested in an appropriate system. Lincomycin was active as inducer in S. lincolnensis, the organism by which it is produced, and streptogramin type B antibiotics induced resistance in S. fradiae, S. hygroscopicus, and the streptogramin type B producer S. diastaticus. Patterns of adenine methylation found included (i) lincomycin-induced monomethylation in S. lincolnensis (and constitutive monomethylation in a mutant selected with maridomycin), (ii) concurrent equimolar levels of adenine mono- plus dimethylation in S. hygroscopicus, (iii) monomethylation in S. fradiae (and dimethylation in a mutant selected with erythromycin), and (iv) adenine dimethylation in S. diastaticus induced by ostreogrycin B.
Mutations in the ilvY gene of Escherichia coli K-12 that cause constitutive expression of ilvC
Authors: Biel A J , Umbarger H E
Published: January 2020

A derivative of Escherichia coli K-12 bearing an ilvC-lac fusion has been studied. beta-Galactosidase formation in this strain is under the control of the ilvC promoter and is therefore induced by the acetohydroxy acids. Derivatives of this fusion strain were isolated that constitutively expressed beta-galactosidase. When an ilvC-containing episome was introduced into these strains, acetohydroxy acid isomeroreductase was also constitutively expressed. The lesions are trans dominant and lie in ilvY, the structural gene specifying a positive control element, v, needed for induction of the isomeroreductase. It was concluded from measurements of beta-galactosidase levels in various diploid strains that, although wild-type v requires inducer to act as a positive control element, it does not act as a repressor in the absence of inducer.
Germination properties of a spore coat-defective mutant of Bacillus subtilis
Authors: Moir A
Published: January 2020

The presence of the gerE36 mutation in strains of Bacillus subtilis 168 resulted in poor germination of their spores in a range of germinants, as measured by the fall in absorbance of spore suspensions. Although resistant to heat and organic solvents, spores were sensitive to lysozyme; electron microscopy revealed that their coat structure was incomplete. These spores responded to germinants by losing heat resistance and changing from phase bright to phase gray. The release of dipicolinic acid and the fall in absorbance of spore suspensions reached only 75 and 50% of wild-type levels, respectively, but followed the same time course as the loss of heat resistance. Although the germination response was incomplete, the concentration of L-alanine required to elicit it was the same for the mutant as for the wild type. The properties of mutant spores suggest that an intact spore coat is not required for the initial interaction between germinant and spore, but that the coat layers may contain molecules important in later stages of germination. In transduction with phage SPP1, the gerE36 mutation mapped between citF and ilvB and was 90% cotransduced with citF2. The gerE mutation identifies the location of a gene important for the progress of late stages of spore formation.
Fractionation studies of the enzyme complex involved in teichoic acid synthesis
Authors: Leaver J , Hancock I C , Baddiley J
Published: January 2020

The membrane-bound enzymes participating in the syntheses of the teichoic acid main chain and linkage unit have been solubilized with Triton X-100 and fractionated by sucrose density gradient centrifugation. Two main fractions were obtained: a heavy fraction, containing enzymes effecting synthesis of the main chain attached to the linkage unit, which was associated with only a small amount of lipid, and a light fraction which was rich in prenyl phosphate and catalyzed only linkage-unit synthesis. The separation by density was not based entirely on polypeptide chain length, as some of the shortest chains appeared in the denser fractions and some relatively high-molecular-weight peptides occurred in the lightest fraction. High activity for linkage-unit synthesis was observed in a fraction containing only a few peptides. Addition of ficaprenyl phosphate to the enzyme preparations had no stimulatory effect. It is concluded that the enzymes for main-chain and linkage unit syntheses frm one or more fairly tightly associated complexes and that polyprenyl phosphate is an integral firmly bound component of the complex in which the linkage unit is synthesized.
Cell wall metabolism in Bacillus subtilis subsp. niger: effects of changes in phosphate supply to the culture
Authors: Kruyssen F J , de Boer W R , Wouters J T
Published: January 2020

Chemostat cultures of Bacillus subtilis subsp. niger WM were exposed to changes in the availability of phosphorus by means of a resuspension technique. Responses in wall metabolism were recorded by measuring the amounts of peptidoglycan and anionic polymers (teichoic or teichuronic acid) in the wall and extracellular fluid fractions. With respect to the wall composition, the effect of a change in orthophosphate supply was a complete shift in the nature of the anionic polymer fraction, the polymer originally present in the walls ("old" polymer) being replaced by the alternative ("new") anionic polymer. The peptidoglycan content of the walls remained constant. It was concluded that the incorporation of old polymer was completely blocked from the moment the orthophosphate supply was changed. However, from a measurement of the total amount of polymer in the whole culture during the course of the experiments, it was evident that synthesis of old polymer continued, but it was secreted. Synthesis of the new polymer started immediately, and it was incorporated exclusively into the wall. During adaption of the cells to the new environment, wall turnover continued in an identical fashion to that extant in steady-state cultures. It was concluded that the primary adaptive response to a change in orthophosphate supply occurred through a mechanism interacting with polymer incorporation and thus at the level of wall assembly at the membrane.
Acid-soluble spore proteins of Bacillus subtilis
Authors: Johnson W C , Tipper D J
Published: January 2020

Acid-soluble spore proteins (ASSPs) comprise about 5% of the total protein of mature spores of different Bacillus subtilis strains. They consist of three abundant species, alpha, beta, and gamma, four less abundant species, and several minor species, alpha, beta, and gamma make up about 18, 18 and 36%, respectively, of the total ASSPs of strain 168, have molecular weights of 5,900, 5,9000, and 11,000, respectively, and resemble the major (A, C, and B) components of Bacillus megaterium ASSPs in several respects, including sensitivity to a specific B. megaterium spore endopeptidase. However, they have pI's of 6.58, 6.67, and 7.96, all lower than those of any of the B. megaterium ASSPs. Although strains varied in the proportions of different ASSPs, to overall patterns seen on gel electrophoresis are constant. ASSPs are located interior to the cortex, presumably in the spore cytoplasm, and are synthesized during sporulation and degraded during germination.
High-frequency chromosome transfer in Rhodopseudomonas sphaeroides promoted by broad-host-range plasmid RP1 carrying mercury transposon Tn501
Authors: Pemberton J M , Bowen A R
Published: January 2020

Insertion of the mercury resistance transposon Tn501 into broad-host-range plasmid RP1 greatly enhanced the ability of this plasmid to promote chromosome transfer in the photosynthetic bacterium Rhodopseudomonas sphaeroides. Compared with the wild-type RP1, which produced less than 10(-8) recombinants per donor cell, RP1::Tn501 produced between 10(-3) and 10(-7) recombinants per donor cell depending upon the marker selected. Plasmid RP1::Tn501 promoted polarized transfer of the chromosome from one or perhaps two origins on the chromosome, giving rise to two linkage groups. All of the biosynthetic and antibiotic resistance genes that have been mapped, including those involved in photosynthesis, occur on one or another of these linkage groups.
Ultrastructure of Naegleria fowleri enflagellation
Authors: Patterson M , Woodworth T W , Marciano-Cabral F , Bradley S G
Published: January 2020

Amoebae of Naegleria fowleri nN68 became elongated flagellated cells 150 to 180 min after subculture to non-nutrient buffer. N. fowleri NF69 did not become elongated or flagellated under these conditions. Electron microscopic examination of N. fowleri confirmed that it is a typical eucaryotic protist with a distinct nuclear envelope and prominent nucleolus, numerous vacuoles and cytoplasmic inclusions, pleomorphic mitochondria, and some rough endoplasmic reticulum. During incubation in non-nutrient buffer, both strains lost ultraviolet-absorbing material to the medium, and the number of vacuoles decreased. In strain nN68, basal bodies, a rootlet, and flagella are formed quickly after an initial lag of 90 min. Initially, the rootlet is not associated with the nucleus but they become associated subsequent at the leading end of the elongated cell. In elongated cells, the rootlet lies in a furrow or groove extending the length of the nucleus. Flagella of N. fowleri nN68 exhibit the typical 9 + 2 arrangement of filaments and are surrounded by a sheath which is continuous with the plasma membrane. The enflagellation process in N. fowleri can be manipulated reproducibly.
Organic nutrition of Beggiatoa sp
Authors: Nelson D C , Castenholz R W
Published: January 2020

Culture OH-75-B of Beggiatoa sp. differed significantly from any described previously in its utilization of organic carbon and reduced sulfur compounds. It deposited internal sulfur granules characteristic of Beggiatoa sp. with either sulfide or thiosulfate in the medium. This strain (OH-75-B, clone 2a) could be grown in agitated liquid cultures on mineral medium with acetate as the only source of organic carbon. The resultant growth yields and rates were comparable to those for typical heterotrophs. Of the other simple organic compounds tested, only pyruvate, lactate, or ethanol could singly support the growth of this strain. Single sugars or amino acids neither supported growth nor enhanced it when added to acetate-containing medium. In contrast, compounds of the tricarboxylic acid cycle enhanced growth yields when tested in concert with acetate. These and fluoroacetate inhibition results indicate that Beggiatoa sp. possesses a functional tricarboxylic acid cycle. Poor yields characterized the growth of this strain on dilute yeast extract medium, and higher concentrations of yeast extract proved inhibitory. The enzyme catalase, contrary to the findings of others, had no synergistic influence on growth yields when added to medium containing yeast extract or acetate or both.
Replication Control and Switch-Off Function as Observed with a Mini-F Factor Plasmid
Authors: Tsutsui Hiroshi , Matsubara Kenichi
Published: January 2020

Mini-F is a fragment of the F plasmid, consisting of 9,000 base pairs, which carries all of the genes and sites required for replicon maintenance and control. Its copy number is one to two per chromosome. This plasmid is joined to ColE1, whose copy number is 16 to 20. Under normal circumstances the composite plasmid replication exhibited ColE1 characteristics, maintaining a high copy number. However, when ColE1 replication was inhibited by deoxyribonucleic acid polymerase I inactivation, its replication exhibited mini-F characteristics, maintaining a low copy number. These observations are in complete agreement with those of Timmis et al. (Proc. Natl. Acad. Sci. U.S.A.
71:
4556-4560, 1974), who examined the behavior of a recombinant plasmid formed between pSC101 and ColE1. The transition from high to low copy number allowed us to examine the control system acting in cells carrying plasmids exhibiting intermediate copy numbers. The initiation of the mini-F replication system as represented by deoxyribonucleic acid synthesis of the composite plasmid was completely blocked when there were multiple copies of mini-F in a cell. It was not restored until the copy number was lowered to one to two, after which replication was first detected. ppF, a mini-F replicon packaged in a phage λ head behaved similarly: its replication was completely shut off when the resident mini-F genome copy number was high and was inhibited partially when the resident mini-F genome copy number was low. These experiments clearly demonstrate that there is a switch-off mechanism acting on deoxyribonucleic acid synthesis (initiation) in a cell carrying mini-F, and its intensity is related to the plasmid copy number. This result supports the “inhibitor dilution model” proposed by Pritchard et al. (Symp. Soc. Gen. Microbiol.
19:
263-297, 1969). The nature of the hypothetical inhibitor is discussed.
Escherichia coli and Salmonella typhimurium supX genes specify deoxyribonucleic acid topoisomerase I
Authors: Trucksis M , Golub E I , Zabel D J , Depew R E
Published: January 2020

Mutations of the Escherichia coli or Salmonella typhimurium supX genes eliminated deoxyribonucleic acid topoisomerase I. Suppression of a supX amber mutation partially restored the topoisomerase. Multicopy plasmids carrying supX+ caused overproduction of topoisomerase. Thus, these supX genes were identified as topA genes which specify deoxyribonucleic acid topoisomerase I.
Effect of Oxygen Radicals and Peroxide on Survival After Ultraviolet Irradiation and Liquid Holding Recovery of
Bacteroides fragilis
Authors: Slade Heide J. K. , Jones David T. , Woods David R.
Published: January 2020

The survival of
Bacteroides fragilis
cells after far-ultraviolet irradiation under anaerobic and aerobic conditions and the liquid holding recovery under aerobic conditions were not affected by peroxide or quenchers of toxic oxygen derivatives.
Complementation of replication-deficient deletion derivatives of plasmid mini-F
Authors: Ebbers J , Eichenlaub R
Published: January 2020

Deleted mini-F plasmids with defects in replication were constructed and tested to see whether they could be rescued through complementation by a helper plasmid. This allowed us to identify two genetic loci determining trans-acting functions required for stable maintenance of plasmid mini-F, one encoded by the PstI fragment from 45.7 to 47.3 F-coordinates (F) and the other most probably located in the region from 43.1 to 43.8 F. The smallest mini-F plasmid that could be established through complementation consists of the PstI fragment 44.0 to 45.7 F, encoding origin II and the incB locus.
High-affinity arabinose transport mutants of Escherichia coli: isolation and gene location
Authors: Clark A F , Hogg R W
Published: January 2020

The gene araF, the product of which is the L-arabinose-binding protein--a component of the high-affinity L-arabinose transport system, was located on the Escherichia coli linkage map at 45 min. We established this location using bacteriophage P2 eductates and bacteriophage P1 cotransduction frequencies with the adjacent genetic loci, his (histidine biosynthesis) and mgl (methylgalactoside transport). In addition, we isolated a number of mutants that phenotypically exhibited altered high-affinity L-arabinose transport capacities. At least two of these mutations were located in the araF gene, as binding protein purified from these strains exhibited altered in vitro arabinose-binding properties.
Restriction map of corynebacteriophages beta c and beta vir and physical localization of the diphtheria tox operon
Authors: Costa J J , Michel J L , Rappuoli R , Murphy J R
Published: January 2020

The BamHI, EcoRI, HindIII, and KpnI restriction endonuclease maps of corynebacteriophage beta c and beta vir were constructed. beta vir appeared to be identical to beta c, except for an approximate 1-kilobase deletion that removed a BamHI site, two KpnI sites, and three EcoRI restriction sites. The diphtheria tox operon was located by hybridizing in vitro 32P-labeled tox messenger ribonucleic acid to blots of endonuclease-digested beta deoxyribonucleic acids. The messenger ribonucleic acid probe was found to hybridize to a 2.1-kilobase region of the beta genome. Since approximately 1.9 kilobases is required to encode prodiphtheria toxin, the data presented strongly suggest that the tox operon of beta is monocistronic.
Cell wall synthesis and initiation of deoxyribonucleic acid replication in Bacillus subtilis
Authors: Sandler N , Keynan A
Published: January 2020

We have observed a connection between cell wall synthesis and the initiation of chromosome replication in Bacillus subtilis. Initiation of chromosome replication was prevented in synchronous cultures in the presence of the cell wall synthesis inhibitor vancomycin. When vancomycin was added to the cultures after initiation of chromosome replication, one round of replication was completed but no reinitiation occurred. Similar results were obtained when cell wall synthesis was inhibited by ristocetin, cycloserine, cloxacillin, or cephaloridine. When sucrose was added to the medium, initiation of deoxyribonucleic acid replication occurred in the presence of vancomycin, to an extent which allowed replication of no more than approximately one-half of the deoxyribonucleic acid of the culture. The same was found in cultures of spheroplasts of B. subtilis. However, initiation of chromosome replication in spheroplasts was completely insensitive to cloxacillin.