bluuebunny/crossref_metadata_2025 huggingface dataset:
Deoxyribonucleic acid and outer membrane: strains diploid for the oriC region show elevated levels of deoxyribonucleic acid-binding protein and evidence for specific binding of the oriC region to outer membrane
Authors: Wolf-Watz H , Masters M
Published: January 2020

We have recently reported that part of the chromosomal deoxyribonucleic acid (DNA) of Escherichia coli is associated with the outer membrane fraction and that an outer membrane protein having a molecular weight of 31,000 probably is involved in this association (H. Wolf-Watz and A. Norqvist, J. Bacteriol. 140:43-49, 1979). We have now found that F' merodiploid strains containing two copies of the DNA between bglB and ilv have increased levels of this protein and an increased amount of DNA in their outer membranes. Increased levels of the protein are also found when lambda asn phage, containing at 1.5-megadalton fragment of DNA located to the right of the uncA uncB genes but to the left of oriC, are induced. It therefore seems that this 1.5-megadalton fragment of DNA either codes for or binds to the 31,000-dalton outer membrane protein. Hybridization studies utilizing DNA found to be bound to outer membrane and DNA isolated from a specialized transducing phage lambda asn 132 revealed that at least 5 to 10% of outer membrane DNA has a DNA sequence homologous with a chromosomal segment carried by this oriC-containing phage.
Macromolecular synthesis in synchronized cultures of Anacystis nidulans
Authors: Asato Y
Published: January 2020

Synchronous culture of Anacystis nidulans has been induced by the light-dark-light regimen. At various time intervals during synchronous growth, samples were pulsed with radioactive labels to determine phospholipid, protein, ribonucleic acid (RNA), and deoxyribonucleic acid (DNA) syntheses within the cell division cycle. A temporal order of protein, RNA, and DNA syntheses occurred within the cell division cycle, whereas phospholipid was characteristically synthesized during midcycle (during cell enlargement) and during the time of cell division. Chemically determined protein, RNA, and DNA syntheses were found to support the schedule of these macromolecules in cultures growing at an 8-h doubling time.
Menaquinone (vitamin K2) biosynthesis: conversion of o-succinylbenzoic acid to 1,4-dihydroxy-2-naphthoic acid by Mycobacterium phlei enzymes
Authors: Meganathan R , Bentley R
Published: January 2020

The coenzyme A (CoA) and adenosine 5'-triphosphate-dependent conversion of o-succinylbenzoic acid (4-[2'-carboxyphenyl]-4-oxobutyric acid) to 1,4-dihydroxy-2-naphthoic acids is an important step in menaquinone (vitamin K2) biosynthesis. Cell-free extracts catalyzing this conversion, obtained from Mycobacterium phlei, were separated into three protein fractions by treatment with protamine sulfate. The second fraction (fraction B) and the supernatant (fraction S) alone did not catalyze dihydroxynaphthoate formation, but did so in combination. All of the results were consistent with the formation of an unstable intermediate, likely an o-succinylbenzoyl-CoA compound, by the action of fraction S. Adenosine 5'-triphosphate was required in this reaction and adenosine 5'-monophosphate was formed. This enzyme activity was termed o-succinylbenzoyl-CoA synthetase: the enzyme showed a marked stability to 0.1 N hydrochloric acid. The presumed o-succinylbenzoyl-CoA derivate was rather unstable; under a variety of conditions, it was converted to a spirodilactone form of o-succinylbenzoate. Fraction B contained an enzyme, termed naphthoate synthase, which converted the o-succinylbenzoyl-CoA derivative to 1,4-dihydroxy-2-naphthoate.
Biosynthesis of glycosylated glycerolphosphate polymers in Streptococcus sanguis
Authors: Mancuso D J , Junker D D , Hsu S C , Chiu T H
Published: January 2020

Two types of glycosylated glycerolphosphates were synthesized when a particulate enzyme prepared from Streptococcus sanguis was incubated with [3H]-phosphatidylglycerol and uridine diphosphate-[14C]glucose in the presence of MgCl2. The first type was extractable with saline and contained no fatty acid. The second type was pellet bound and could be extracted with 0.1% sodium dodecyl sulfate. Both types of polymers were purified and partially characterized. The first type of polymer was fractionated into five polymers, peaks 2a, 2b, 2c, 3a, and 3b. All except peak 2a, which contained only [3H]glycerol, contained both [3H]glycerol and [14C]glucose. [3H]NaBH4 reduction of acid hydrolysates of the polymers revealed that all of the polymers contained glucose as the major sugar componenta nd xylose as the minor sugar component. The second type of polymer was fractionated into three polymers, P-1, P-2, and P-3. All contained [3H]-glycerol, [14C]glucose, and fatty acids. P-1 appeared to be pure, whereas P-2 and P-3 contained two polymers each, as judged from sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Caulobacter cresentus mutant defective in membrane phospholipid synthesis
Authors: Contreras I , Bender R A , Mansour J , Henry S , Shapiro L
Published: January 2020

To study the relationship between phospholipid synthesis and organelle biogenesis in the dimorphic bacterium Caulobacter crescentus, auxotrophs have been isolated which require exogenous glycerol or glycerol 3-phosphate for growth when glucose is used as the carbon source. Upon glycerol deprivation, net phospholipid synthesis ceased immediately in a glycerol 3-phosphate auxotroph which was shown to have levels of biosynthetic sn-glycerol 3-phosphate dehydrogenase (E.C. 1.1.1.8) activity 10 times lower than that of the wild type. In the absence of glycerol, the optical density of the culture continued to increase for the equivalent of one generation, although the cells did not divide. After the equivalent of one generation time, rapid cell death occurred. Cell death also occurred when phospholipid synthesis was inhibited by cerulenin. Although ribonucleic acid and protein syntheses continued at a reduced rate for the equivalent of one generation in mutant strains, a substantial decrease in the rate of deoxyribonucleic acid synthesis occurred immediately upon glycerol deprivation. Revertant strains had wild-type levels of glycerol 3-phosphate dehydrogenase activity and normal rates of phospholipid and macromolecular synthesis.
Existence of the bound form of prolipoprotein in Escherichia coli B cells treated with globomycin
Authors: Inukai M , Takeuchi M , Shimizu K , Arai M
Published: January 2020

A murein-bound form of prolipoprotein was found in the cell envelope fraction of globomycin-treated Escherichia coli B. We suggest therefore that proteolytic cleavage of prolipoprotein to mature lipoprotein is not essential for the transpeptidation of the lipoprotein to peptidoglycan.
Direction of deoxyribonucleic acid transfer and replication in a derivative of plasmid R100-1
Authors: Legrand P , Bouche J P , Louarn J M
Published: January 2020

The site of integration and the molecular orientation of a prophage Mu integrated within the resistance transfer factor component of plasmid R100-1 have been determined on the physical map of the plasmid. This allowed us (i) to determine the direction of deoxyribonucleic acid transfer from oriT during conjugation and (ii) to demonstrate the unidirectionality of replication in conditions of exponential growth (by determining the strand preference of Mu-specific Okazaki fragments).
Hybrid tryptophan synthase beta 2 proteins: apparent conservation of the beta-beta binding region of the beta monomer among enteric bacteria
Authors: Brennan E F , Rocha V
Published: January 2020

Purified borohydride-reduced tryptophan synthase beta 2 protein (EC 4.2.1.20) from Escherichia coli and purified native beta 2 protein from Serratia marcescens were mixed and dissociated in urea. Removal of the urea resulted in random reassociation of the reduced and native beta monomers, forming interspecies hybrid beta 2 molecules. Interspecies hybrid beta 2 protein molecules of the reciprocal composition were also formed. Interspecies hybrid reconstituted molecules were formed with approximately the same efficiency as intraspecies reconstituted molecules (reduced and native monomers from the same species) indicating no particular preference for reassembly. The data provide evidence that the structural region of interaction between the beta monomers necessary for dimerization is highly conserved in the enzymes from the two organisms examined.
Lipolytic activity copurified with the outer membrane of Serratia marcescens
Authors: Heller K B
Published: January 2020

Lipase, nuclease, and protease activities could be shown primarily with the purified outer membrane fraction from Serratia marcescens. These activities increased and decreased in the different compartments dependent on the growth phase of the cell culture. Penicillin-hydrolyzing activity was exclusively demonstrated with the outer membrane fraction.
Siderophore synthesis in Klebsiella pneumoniae and Shigella sonnei during iron deficiency
Authors: Perry R D , San Clemente C L
Published: January 2020

Klebsiella pneumoniae 298/53 and Shigella sonnei 43-GG9 exhibited restricted growth and enterochelin synthesis only under iron-deficient conditions. S. sonnei also produced an unidentified secondary hydroxamate siderophore.
Separation and partial characterization of two deoxyribonucleic acid polymerases from Spiroplasma citri
Authors: Charron A , Bebear C , Brun G , Yot P , Latrille J , Bove J M
Published: January 2020

The separation and partial characterization of two deoxyribonucleic acid polymerases from Spiroplasma citri have been achieved. The two enzymes had different elution properties on diethylaminoethyl (DEAE) cellulose and differed in their sensitivity to N-ethylmaleimide (NEM), preference for different template-primers, and sedimentation velocity in linear glycerol gradients. The first enzyme activity, ScA, was retained on DEAE-cellulose and was not inhibited by NEM. Activated deoxyribonucleic acid and poly(dA)-oligo(dT12) were the preferred template-primers. Arabinosyl-cytidine triphosphate had no effect. The sedimentation coefficient of ScA was 6.3s. The second activity, ScB, was not retained on DEAE-cellulose and was inhibited by NEM. Poly(dA)-oligo(dT12) was the preferred template-primer, whereas activated DNA was only poorly utilized. ScB was not affected by arabinosyl-cytidine triphosphate, and its sedimentation coefficient was 4.4s. The polymerization activities of the two enzymes were maximum at 37 to 40 degrees C.
Response to a metal ion-citrate complex in bacterial sensing
Authors: Ingolia T D , Koshland D E
Published: January 2020

Salmonella typhimurium responds chemotactically to gradients of divalent cations in the presence of citrate ions. The actual chemoeffector is the citrate-metal ion complex, which acts as an attractant. Citrate (which is also a chemoeffector for Salmonella) and the citrate-metal ion complex are recognized by different receptors. The response of Salmonells, which can transport citrate through its membrane, is quite different than that of Escherichia coli, which cannot.
Binding of yeast killer toxin to a cell wall receptor on sensitive Saccharomyces cerevisiae
Authors: Bussey H , Saville D , Hutchins K , Palfree R G
Published: January 2020

35S-labeled killer toxin protein bound to cells of sensitive Saccharomyces cerevisiae S14a. Strains that were resistant to toxin through mutation in the nuclear genes kre1 kre2 bound toxin only weakly. Non-radioactive toxin competed effectively with 35S-labeled toxin for binding to S14a, but did not compete significantly in the binding to mutant kre1-1. This implied that binding to kre1-1 was nonspecific. A Scatchard analysis of the specific binding to S14a gave a linear plot, with an association constant of 2.9 x 10(6) M-1 and a receptor number of 1.1 x 10(7) per cell. Killer toxin receptors were solubilized from the cell wall by zymolyase digestion. Soluble, non-dialyzable cell wall digest from S14a competed with sensitive yeast cells for 35S-labeled toxin binding and reduced toxin-dependent killing of a sensitive strain. Wall digest from kre1-1 competed only weakly for toxin binding with sensitive cells and caused little reduction of toxin-dependent killing. Although the abundant (1.1 x 10(7) per cell) receptor appeared necessary for toxin action, as few as 2.8 x 10(4) toxin molecules were necessary to kill a sensitive cell of S14a. The kinetics killing of S14a suggested that some component was saturated with toxin at a concentration 50-fold lower than that needed to saturate the wall receptor.
Isolation and characterization of an R-prime plasmid from Rhizobium meliloti
Authors: Kiss G B , Dobo K , Dusha I , Breznovits A , Orosz L , Vincze E , Kondorosi A
Published: January 2020

Using a simple enrichment procedure, we isolated an R-prime derivative of plasmid R68.45 carrying a 17.8-megadalton segment of the Rhizobium meliloti 41 chromosome. The chromosomal segment carried on this plasmid (pGY1) includes the markers cys-24+, cys-46+, and att16-3. Plasmid pGY1 mobilized the chromosome in a polarized way starting from the region of homology, but cannot promote chromosome transfer from other sites. The att16-3 site on pGY1 allowed the integration of phage 16-3 into pGY1, and a composite plasmid of 91.8 megadaltons was formed. This vector (pGY2) is suitable for the introduction of Rhizobium bacteriophage 16-3 into other gram-negative bacteria.
Escherichia coli regulatory mutation affecting lysine transport and lysine decarboxylase
Authors: Popkin P S , Maas W K
Published: January 2020

A spontaneous thiosine-resistant mutant of Escherichia coli was shown to have the following characteristics: lowered initial rate of lysine uptake and lowered plateau level of accumulation of exogenous lysine by both the lysine-specific and the general basic amino acid transport systems; altered repressibility of these two lysine transport systems; a derepressed level of lysine decarboxylase; normal growth rate; parental levels of lysyl-transfer ribonucleic acid synthetase and the inducible and constitutive arginine and ornithine decarboxylases. Both the mutant (lysP) and its parent (lysP+) feed a lysine auxotroph when they are plated in proximity on solid medium. However, the feeding response was observable after 1 day less of incubation when the mutant was the feeding strain. Despite the derepressed level of lysine decarboxylase in exponential cultures of the mutant extracts of these cultures had no detectable cadaverine pool. Conjugation experiments established the following gene order: gyrA (formerly nalA) lysP metG his. All thiosine-resistant recombinants assayed showed reduced lysine transport. In many of these recombinants the derepression of lysine decarboxylase was not expressed.
Assembly of outer membrane lipoprotein in an Escherichia coli mutant with a single amino acid replacement within the signal sequence of prolipoprotein
Authors: Lin J J , Kanazawa H , Wu H C
Published: January 2020

We have compared the rate of assembly of outer membrane proteins including the lipoprotein in a pair of isogenic mlpA+ (lpp+) and mlpA (lpp) strains by pulse-chase experiments. The rate of assembly of the mutant prolipoprotein into the outer membrane was slightly slower than that of the wild-type lipoprotein. The rate of assembly of protein I and protein H-2 was similar in the wild type and the mutant, whereas the rate of assembly of protein II into the outer membrane was slightly reduced in the mutant strain. The organization of outer membrane was slightly reduced in the mutant strain. The organization of outer membrane proteins in the mutant cells appeared not to be grossly altered, based on the apparent resistance (or susceptibility) of these proteins toward trypsin treatment and their resistance to solubilization by Sarkosyl. Like the wild-type lipoprotein, the mutant prolipoprotein in the outer membrane was resistant to trypsin. On the other hand, the prolipoprotein in the cytoplasmic membrane fraction of the mutant cell envelope was susceptible to trypsin digestion. We conclude from these data that proteolytic cleavage of prolipoprotein is not essential for the translocation and proper assembly of lipoprotein into outer membrane.
Regulation of cholera toxin production in Vibrio cholerae: genetic analysis of phenotypic instability in hypertoxinogenic mutants
Authors: Mekalanos J J , Murphy J R
Published: January 2020

Hypertoxinogenic mutants of Vibrio cholerae frequently possess mutations in a chromosomal locus called htx. Spontaneously occurring phenotypic revertants were shown to fall into three classes. One class retained the htx mutation and therefore represented a second-site mutation(s) capable of suppressing the Htx phenotype, whereas the other two classes represented strains that had lost the htx mutation. One of the latter two classes appeared to be composed of true genetic revertants, whereas the third class consisted of clones that had replaced htx with a new mutation conferring a hypotoxinogenic phenotype. Several rare N-methyl-N'-nitro-N-nitrosoguanidine-induced hypotoxinogenic mutants selected by rif comutation also map in the same region as htx. This new locus, which mediates the hypotoxinogenic phenotype in these trains and potentially in some hypertoxinogenic phenotypic revertants, has been designated ltx. htx and ltx appear to be regulatory loci, since mutations in both sites alter the level of cholera toxin A and B subunit production coordinately. The genetic data also support a model in which htx- and ltx- are allelic states of the same Tox regulatory locus.
Isolation and Characterization of a Composite Plasmid Rms201 Mutant Temperature Sensitive for Replication
Authors: Ike Yasuyoshi , Hashimoto Hajime , Motohashi Kazuo , Fujisawa Noriko , Mitsuhashi Susumu
Published: January 2020

A mutant temperature-sensitive for R-plasmid replication, Rms201
ts
14, was isolated from composite plasmid Rms201 after mutagenesis of P1 transducing lysate with 100 mM hydroxylamine for 40 h at 37°C. When
Escherichia coli
ML1410(Rms201
ts
14)
+
was grown at temperatures between 40 and 42°C in L broth, antibiotic-sensitive cells were segregated. When the incubation temperature of ML1410(Rms201
ts
14)
+
in L-broth was shifted to 42 from 30°C, the increase in the number of antibiotic-resistant cells ceased 90 min after the temperature shift. However, the total number of cells continuously increased, and only 3% of the cells retained the plasmid at 5 h after the temperature shift to 42°C. At 30°C the amounts of covalently closed circular deoxyribonucleic acid per chromosome of Rms201
ts
14 and Rms201 were 3.8 and 6.3%, respectively. Incorporation of radioactive thymidine into the covalently closed circular deoxyribonucleic acid of Rms201
ts
14 did not take place at 42°C, whereas radioactive thymidine was incorporated into the covalently closed circular deoxyribonucleic acid of Rms201 at a rate of 4%/chromosome even at 42°C. The synthesis of plasmid covalently closed circular deoxyribonucleic acid in a cell harboring Rms201
ts
14 was almost completely blocked at 42°C. These results indicated that the gene(s) responsible for plasmid deoxyribonucleic acid replication was affected in the mutant Rms201
ts
14. Temperature-sensitive miniplasmid pMS
ts
214, which has a molecular weight of 5.3 × 10
6
and encodes ampicillin resistance, was isolated from Rms201
ts
14. Similarly, miniplasmid pMS201, which encodes single ampicillin resistance, was isolated from its parent, Rms201, and its molecular weight was 4.7 × 10
6
. These results indicate that the gene(s) causing temperature sensitivity for replication of Rms201 resides on the miniplasmid.
Behavioral mutants of Rhizobium meliloti
Authors: Ames P , Schluederberg S A , Bergman K
Published: January 2020

Rhizobium meliloti exhibited behavior resembling that of other chemotactic bacteria. Nonflagellated, nonmotile but flagellated, and nonchemotactic mutants were isolated, and genetic mapping placed all three types of lesions within a small region of the R. Meliloti chromosome. Results from nodulation experiments performed with these mutants indicate that members of the three classes (Fla-, Mot-, and Che-) infect and produce as many nodules on alfalfa plants as does the wild-type strain.
Methyl-coenzyme M, an intermediate in methanogenic dissimilation of C1 compounds by Methanosarcina barkeri
Authors: Shapiro S , Wolfe R S
Published: January 2020

Extracts of Methanosarcina barkeri possess a specific methyltransferase that catalyzes the transfer of the methyl group of methanol to 2-mercaptoethanesulfonic acid. Over a fourfold range in added 2-mercaptoethanesulfonic acid, the formation of 2-(methylthio)ethanesulfonic acid exhibited a 1:1 ratio to 2-mercaptoethanesulfonic acid added. This reaction required adenosine 5'-triphosphate; a maximal ratio (mole/mole) of 85 methyl groups was transferred per adenosine 5'-triphosphate added. The methyltransferase was found in extracts of methanol-grown cells as well as in extracts of hydrogen-grown cells. In extracts of cells grown on either substrate, 2-(methylthio)ethanesulfonic acid was formed from added methanol or methylamine but not from acetate.
Ionic control of germination of Blastocladiella emersonii zoospores
Authors: Van Brunt J , Harold F M
Published: January 2020

Encystment and germination of Blastocladiella emersonii zoospores involve a rapid and radical transformation of the motile but nongrowing spore into a sessile, growing germling. Certain inorganic ions, notably 50 mM KCl, are efficient inducers of germination. By use of the carbocyanine dye DiO-C6-(3), we found that KCl depolarizes the plasma membrane of zoospores and noted good correlation between depolarization and subsequent germination. Zoospores avidly accumulated K+ ions from the medium, attaining an internal concentration of over 50 mM and a concentration gradient of 2,500. Sodium ions, by contrast, were expelled. Internal K+ was required for normal germination but its function is not known. Zoospores also took up considerable amounts of calcium; most of this was associated with the external surface and appeared to be necessary for maintenance of zoospore integrity. KCl (50 mM) and other salts displaced surface calcium but this was not in itself sufficient to induce germination. The calcium ionophore A23187, in the presence of external calcium, was an effective inducer of germination, suggesting a possible role for cytosolic calcium in triggering the transformation. We propose that the first step in the induction of germination by salts is depolarization of the plasma membrane; subsequent events require the intervention of cytoplasmic signals.
Subcellular distribution of marker enzymes in cells of a minute fungus, Fusidium sp. 100-3
Authors: Handley D , Ghosh B K
Published: January 2020

An electron microscope cytochemical technique was used to determine the subcellular distribution of marker enzymes in Fusidium sp. 100-3 cells. Nucleoside diphosphatase was found in the nuclear envelope and intracytoplasmic membrane segment. Thiamine pyrophosphatase was found to be associated with the mesosomes. Cytochrome c (oxidase) activity was found only in the mitochondrial cristae. Strong alkaline phosphatase activity was present in the vacuole; in addition, the enzyme activity was discretely dispersed throughout the cytoplasm without any association with any membrane material. The overall characteristics of the cell ultrastructure and subcellular enzyme distribution of Fusidium sp. 100-3 cells compare fairly well with those of a fungal cell. But there are considerable differences from the characteristics of higher eucaryotic cells. Detailed data on the marker enzymes distribution in a variety of fungal cells are not available. Therefore, it is not possible to conclude whether the marker enzyme distribution of Fusidium sp. 100-3 cells is unique or is typical of any fungal organism. Detailed studies of cell ultrastructure of and marker enzyme distribution in minute fungal cells and their comparison to the ultrastructure of and marker enzyme distribution in other fungal organisms may be helpful in understanding the phylogenetic and ontogenic development of subcellular organelles.
Regulatory effect of hydrogen on nitrogenase activity of the blue-green alga (cyanobacterium) Nostoc muscorum
Authors: Scherer S , Kerfin W , Böger P
Published: January 2020

Preincubation of the blue-green alga (cyanobacterium) Nostoc muscorum under an atmosphere of argon plus acetylene in the light led to a greater than fourfold increase of light-induced hydrogen evolution and to a 50% increase of acetylene reduction, as compared to cells that had not been preconditioned. The basic and the increased hydrogen evolution were both due to nitrogenase activity. Furthermore, after preincubation the hydrogen uptake, usually observed with unconditional cells, was abolished. Nostoc preincubated under acetylene evolved hydrogen in the light even in the presence of nitrogen for at least 2 h, with a 15-fold increase as compared to the unconditioned cells. These acetylene effects could be completely abolished by the presence of hydrogen during acetylene preincubation. These findings indicate that the hydrogen concentration in N. muscorum cells plays a role in regulation of nitrogenase activity.
Genetic and physiological characterization of Escherichia coli mutants deficient in phosphoenolpyruvate carboxykinase activity
Authors: Goldie A H , Sanwal B D
Published: January 2020

Mutants doubly deficient in phosphoenolpyruvate carboxykinase (pck) and phosphoenolpyruvate synthetase (pps) were unable to grow with succinate as the sole carbon source. A number of pck mutations isolated from pps strains by penicillin selection mapped at 74 min on the Escherichia coli chromosome, between glpD and aroB. Several of the strains containing these mutations had a protein antigenically related to phosphoenolpyruvate carboxykinase, and therefore, the mutations probably represented mutations in the structural gene for this enzyme. Phosphoenolpyruvate carboxykinase was regulated at the level of transcription by catabolite repression. Enzyme levels also increased in stationary-phase cultures by a mechanism independent of cyclic adenosine monophosphate or the product of the relA gene.
Diversity among B6 strains of Agrobacterium tumefaciens
Authors: Hamada S E , Farrand S K
Published: January 2020

A total of 20 laboratory substrains of Agrobacterium tumefaciens strain B6 were compared with respect to six characteristics, including 3-ketolactose production, lysogeny, octopine catabolism, tumorigenic host range, and plasmid content. Within this group of strains diversity was found for all characteristics except 3-ketolactose production. Six substrains were lysogenized with an omega-type phage, whereas one substrain appeared neither sensitive to nor lysogenized with this bacteriophage. All but two substrains catabolized octopine and induced tumors on carrot disks. These 18 substrains harbor deoxyribonucleic acid sequences homologous to pTiB6-806. The two substrains unable to catabolize octopine were nontumorigenic and lacked detectable Ti plasmid sequences. Of the 20 substrains, 13 also contained sequences homologous to the cryptic plasmid pAtB6-806; 2 of the 18 substrains tumorigenic on carrots failed to induce tumors on Kalanchoe leaves. Their inability to induced tumors on this host, could not be correlated with lysogeny, with the presence or absence of pAtB6-806, or with the very large cryptic plasmid recently described. The Ti plasmids from these two strains were indistinguishable from pTiB6-806 by restriction enzyme analysis and could genetically convert a cured A. tumefaciens strain to tumorigenicity on both plant species. The results with these two strains suggest that parameters of tumorigenicity, such as host range, may be controlled by the bacterial chromosome.
Isolation and characterization of lambda specialized transducing bacteriophages carrying Klebsiella pneumoniae nif genes
Authors: MacNeil D , Howe M M , Brill W J
Published: January 2020

Seve lambda dnif specialized transducing bacteriophages were isolated from Escherichia coli strains containing plasmids carrying the his-nif region of Klebsiella pneumoniae. These phages collectively carry deoxyribonucleic acid for all of the genes in the nif regulon and adjacent deoxyribonucleic acid of K. pneumoniae. The phages were isolated by using Mu insertions in the nif region to direct the integration of lambda pMu phages in nif via formation of lambda pMu-Mu dilysogens which, upon induction, yielded lambda dnif phages. This procedure should be generally applicable for isolating lambda specialized transducing phages carrying genes from E. coli or other bacteria.
Interaction of mutagenic spermidine-nitrous acid reaction products with uvr- and recA-dependent repair systems in Salmonella
Authors: Murphey-Corb M , Kong H L , Murray M L
Published: January 2020

It has been observed previously that the mutagenic action of nitrous acid may be potentiated by polyamines. We examined the cellular response of two deoxyribonucleic acid repair systems to treatment with spermidine-nitrite reaction products. uvrB- deficient mutants of Salmonella typhimurium LT2 showed enhanced lethal and mutagenic response to the reaction products. Lethal activity was further enhanced in a uvrB recA double mutant, whereas mutagenic activity was not detectable. Dependence of mutagenesis on the recA gene implicates the action of an error-prone repair system in the fixation of a premutagenic lesion as a mutation. From consideration of the substrate characteristics of the two repair systems studied, it is suggested that the deoxyribonucleic acid lesion formed by the reaction products of spermidine and nitrite is an intrastrand cross-link.
S-adenosylmethionine levels and protein methylation during morphogenesis of Mucor racemosus
Authors: Garcia J R , Hiatt W R , Peters J , Sypherd P S
Published: January 2020

The intracellular level of S-adenosylmethionine increased as the yeast-phase cells of Mucor racemosus were induced to convert to hyphae. This increase correlated well with time course of the conversion in cell type and was independent of the metabolic changes caused by the shift to aerobic conditions. There was no significant change in the intracellular level of spermidine, a polyamine synthesized from putrescine and the propylamine group of S-adenosylmethionine. Spermine was not detected. An examination of protein methylation revealed an increase in the methylation of total protein during the shift in cell type and possible qualitative as well as quantitative changes in specific base proteins.
Ultraviolet-endonuclease activity in cell extracts of Saccharomyces cerevisiae mutants defective in excision of pyrimidine dimers
Authors: Bekker M L , Kaboev O K , Akhmedov A T , Luchkina L A
Published: January 2020

Cell-free extracts of ultraviolet-sensitive mutants of Saccharomyces cerevisiae defective in excision of pyrimidine dimers, rad1, rad2, rad3, rad4, rad10, and rad16, as well as the extracts of the wild-type strain RAD+, display ultraviolet-endonuclease activity.
Purification of glutamine synthetase from a variety of bacteria
Authors: Streicher S L , Tyler B
Published: January 2020

We have developed two procedures which allow the very rapid purification of glutamine synthetase (GS) from a diverse variety of bacteria. The first procedure, based upon differential sedimentation, depends upon the association of GS with deoxyribonucleic acid in cell extracts. The second procedure, derived from the method of C. Gross et al (J. Bacteriol. 128:382-389, 1976) for purifying ribonucleic acid polymerase by polyethylene glycol (PEG) precipitation, enabled us to obtain high yields of GS from either small or large quantities of cells. We used the PEG procedure to purify GS from Klebsiella aerogenes, K. pneumoniae, Escherichia coli, Salmonella typhimurium, Rhizobium sp. strain 32H1, R. meliloti, Azotobacter vinelandii, Pseudomonas putida, Caulobacter crescentus, and Rhodopseudomonas capsulata. The purity of the GS obtained, judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was high, and in many instances only a single protein band was detected.
Mutations which affect the structure and activity of colicin E3
Authors: Mock M , Schwartz M
Published: January 2020

Among 69 ColE3 mutant plasmids selected on the basis of their inability to produce an active colicin, seven (cop-1 to cop-7) were found to bear a mutation affecting the structural gene for colicin. Three of these (cop-1, cop-2 and cop-3) lead to the production of an inactive colicin molecule which has the same molecular weight as wild-type colicin E3 (67,000). These three inactive colicins are still able to interact with the outer membrane receptor. The cop-1 protein retained the ability to inhibit protein synthesis in vitro and therefore seems specifically affected in it ability to penetrate the cell envelope. The cop-2 and cop-3 proteins lost the ability to inhibit protein synthesis in vitro, and activity which is normally associated with the C-terminal part of the colicin molecule. On the basis of this and further evidence, it is suggested that the cop-2 and cop-3 mutations affect the structure of the C-terminal part of the colicin molecule. The other four mutations (cop-4 to cop-7) lead to the production of colicin-related polypeptides of lower molecular weight (29,000 to 45,000) which display a reaction of partial immunological identity with wild-type colicin. These four polypeptides are unable to interact with the cell surface receptor. Three of these mutants are shown to carry a nonsense mutation.
Transferability of macrolide, lincomycin, and streptogramin resistances between group A, B, and D streptococci, Streptococcus pneumoniae, and Staphylococcus aureus
Authors: Engel H W , Soedirman N , Rost J A , van Leeuwen W J , van Embden J D
Published: January 2020

The transferability of plasmid pRI405 between various streptococci of groups A, B, and D, Streptococcus pneumoniae, and Staphylococcus aureus is described. pRI405 originated from Streptococcus faecalis and encodes for resistance to macrolides, lincomycin, and streptogramin B (MLS resistance). The host range of the well-documented streptococcal plasmid pAM beta 1 was found to be similar to that of pRI405. Cleavage with restriction enzymes suggests that pRI405 belongs to a related family of MLS resistance plasmids.
Construction and Characterization of
Salmonella typhimurium
Strains That Accumulate and Excrete α- and β- Isopropylmalate
Authors: Fultz Patricia N. , Choung Kyung K. L. , Kemper Jost
Published: January 2020

Two
Salmonella typhimurium
strains, which could be used as sources for the leucine biosynthetic intermediates α- and β-isopropylmalate were constructed by a series of P22-mediated transductions. One strain, JK527 [
flr-19 leuA2010
Δ(
leuD-ara
)
798 fol-162
], accumulated and excreted α-isopropylmalate, whereas the second strain, JK553 (
flr-19 leuA2010 leuB698
), accumulated and excreted α- and β-isopropylmalate. The yield of α-isopropylmalate isolated from the culture medium of JK527 was more than five times the amount obtained from a comparable volume of medium in which
Neurospora crassa
strain FLR
92
-1-216 (normally used as the source for α- and β-isopropylmalate) was grown. Not only was the yield greater, but
S. typhimurium
strains are much easier to handle and grow to saturation much faster than
N. crassa
strains. The combination of the two regulatory mutations
flr-19
, which results in constitutive expression of the leucine operon, and
leuA2010
, which renders the first leucine-specific biosynthetic enzyme insensitive to feedback inhibition by leucine, generated limitations in the production of valine and pantothenic acid. The efficient, irreversible, and unregulated conversion of α-ketoisovaleric acid into α-isopropylmalate (α-isopropylmalate synthetase

K
m

for α-ketoisovaleric acid, 6 × 10
−5
M) severely restricted the amount of α-ketoisovaleric acid available for conversion into valine and pantothenic acid (ketopantoate hydroxymethyltransferase

K
m

for α-ketoisovaleric acid, 1.1 × 10
−3
M; transaminase B

K
m

for α-ketoisovaleric acid, 2 × 10
−3
M).
Stimulation of yeast ascospore germination and outgrowth by S-adenosylmethionine
Authors: Brawley J V , Ferro A J
Published: January 2020

The supplementation of S-adenosylmethionine (SAM) to germination medium stimulated the accumulation of [14C]uracil from the medium into germinating cells, as well as its incorporation into ribonucleic acid during germination and outgrowth of ascospores of Saccharomyces cerevisiae. In addition to uracil, the accumulation of leucine, cytosine, serine, and methionine was also stimulated by the extracellular addition of this sulfonium compound. The SAM-stimulatory effect was dose dependent; half-maximal stimulation was observed at about 50 muM. The effect exerted by SAM supplementation appeared to be specific for SAM and for germination and outgrowth. In the absence of SAM biosynthesis (in the presence of cycloleucine), spores were inhibited in their ability to accumulate label, whereas the supplementation of SAM completely reversed the cycloleucine-induced inhibition of accumulation. In addition to accumulation and incorporation, the kinetics of bud formation during outgrowth were also stimulated by exogenous SAM. The stimulation of budding by SAM was amplified in an ethionine-resistant strain. These observations suggest that SAM may be essential for the initiation of cell division during the breaking of spore dormancy.
Regulation of Glucose Transport and Metabolism in
Thiobacillus novellus
Authors: Matin A. , Schleiss M. , Perez R. C.
Published: January 2020

To investigate the physiological basis of decreased rate of glucose utilization by
Thiobacillus novellus
in a mixotrophic environment (R. C. Perez and A. Matin, J. Bacteriol.
142:
633–638, 1980), its glucose transport system was characterized and the modulation of this system as well as enzymes of glucose metabolism by the growth environment was examined. Uptake of 2-deoxy-
d
-glucose by cell suspensions was almost abolished by respiratory chain inhibitors, and the sugar accumulated unchanged inside the cells against a concentration gradient: its transport is probably linked to the proton electrochemical gradient. The glucose transport system, as well as several enzymes of glucose metabolism, had a high specific activity in heterotrophic cells, intermediate activity in mixotrophic cells, and low activity in autotrophic cells; thus, they are induced by glucose but repressed by thiosulfate, its metabolites, or both. Thiosulfate and sulfite inhibited the glucose transport system uncompetitively and noncompetitively, respectively (apparent

K
i

= 3.1 × 10
−2
M and 3.3 × 10
−7
M, respectively) and also inhibited glucose-6-phosphate dehydrogenase activity. Thus, the rate of glucose utilization in mixotrophic environments decreased because thiosulfate and its metabolites repress as well as inhibit the glucose transport system and enzymes of glucose metabolism. The significance of this and other regulatory phenomena that come into play in such environments is discussed.
Intramolecular transposition and inversion in plasmid R6K
Authors: Chiang S J , Clowes R C
Published: January 2020

Selection was made in Escherichia coli K-12 recA hosts carrying plasmid R6K for ampicillin hyperresistance. Twenty-two selected strains were found to carry mutant plasmids, which, from electron microscopy and restriction enzyme analysis, were concluded to arise by a duplication of transposon Tn2660, which confers ampicillin resistance, in all cases the duplicate transposon being in an inverted orientation with respect to the resident Tn2660. A mutant of R6K, pSJC301, which was temperature sensitive for ampicillin resistance was produced by in vitro hydroxylamine treatment of R6K deoxyribonucleic acid. A plasmid hybrid, pSJC102, was constructed by cloning the EcoRI R6K fragment carrying the wild-type beta-lactamase gene into the EcoRI site of ColE1. pSJC301 and pSJC102 were transformed into the same recA host strain to form a stable biplasmid strain. Ampicillin-hyperresistant mutants were selected from this strain and screened for plasmids with a duplication of transposon Tn2660, which occurred with equal frequency in either pSJC301 or pSJC102; of 12 characterized, all were inverse repeats of the resident transposon. All six Tn2660 inserts into pSJC301 determined temperature-sensitive ampicillin resistance, and all six inserts into pSJC102 determined wild-type ampicillin resistance, from which it was inferred that transposition of a duplicate Tn2660 occurs predominantly as an intramolecular event, at least in the multicopy R6K plasmid. In all 28 insertion mutants of R6K, there was an inversion of the deoxyribonucleic acid between the two transposons, whereas in only one of six insertion mutants of pSJC102, inversion had occurred. These results are discussed in terms of current models of transposition.
Gene ilvY of Salmonella typhimurium
Authors: Blazey D L , Burns R O
Published: January 2020

Evidence is presented for the existence in Salmonella typhimurium LT2 of the regulatory gene ilv Y. The Escherichia coli K-12 ilvY gene product is shown to complement a S. typhimurium ilvY mutation in vivo.
A mutant Ebg enzyme that converts lactose into an inducer of the lac operon
Authors: Rolseth S J , Fried V A , Hall B G
Published: January 2020

Lactose is not itself an inducer of the lac operon, nor is it converted to an inducer by ebg+ beta-galactosidase of Escherichia coli. We report here the isolation of a mutant Ebg beta-galactosidase which is capable of converting lactose into an inducer of the lac operon.
Physiological Studies of β-Galactosidase Induction in
Kluyveromyces lactis
Authors: Dickson Robert C. , Markin Jennifer S.
Published: January 2020

We examined the kinetics of β-galactosidase (EC 3.2.1.23) induction in the yeast
Kluyveromyces lactis
. Enzyme activity began to increase 10 to 15 min, about 1/10 of a cell generation, after the addition of inducer and continued to increase linearly for from 7 to 9 cell generations before reaching a maximum, some 125- to 150-fold above the basal level of uninduced cells. Thereafter, as long as logarithmic growth was maintained, enzyme levels remained high, but enzyme levels dropped to a value only 5- to 10-fold above the basal level if cells entered stationary phase. Enzyme induction required the constant presence of inducer, since removal of inducer caused a reduction in enzyme level. Three nongratuitous inducers of β-galactosidase activity, lactose, galactose, and lactobionic acid, were identified. Several inducers of the
lac
operon of
Escherichia coli
, including methyl-, isopropyl- and phenyl-1-thio-β-
d
-galactoside, and thioallolactose did not induce β-galactosidase in
K. lactis
even though they entered the cell. The maximum rate of enzyme induction was only achieved with lactose concentrations of greater than 1 to 2 mM. The initial differential rate of β-galactosidase appearance after induction was reduced in medium containing glucose, indicating transient carbon catabolite repression. However, glucose did not exclude lactose from
K. lactis
, it did not cause permanent carbon catabolite repression of β-galactosidase synthesis, and it did not prevent lactose utilization. These three results are in direct contrast to those observed for lactose utilization in
E. coli
. Furthermore, these results, along with our observation that
K. lactis
grew slightly faster on lactose than on glucose, indicate that this organism has evolved an efficient system for utilizing lactose.
Transport of methylamine by Pseudomonas sp. MA
Authors: Bellion E , Khan M Y , Romano M J
Published: January 2020

Pseudomonas sp. MA grows on methylamines as a sole source of carbon, nitrogen, and energy. The transport of methylamine into the organism was investigated. It was found that this organism possesses an inducible transport system for methylamine having the following physical parameters: pH optimum, 7.2; temperature optimum, 30 to 35 degrees C; Km, 1 to 30 mM; Vmax, 90 to 120 nmol/min per mg (dry weight) of cells. Methylamine uptake was curtailed by azide, cyanide, and carbonyl cyanide-m-chlorophenylhydrazone; osmotic shock treatment reduced the uptake by 50%. The uptake was not effectively inhibited by ammonium ion, amino acids, or amides, but was competitively inhibited by short-chain alkylamines. Cells grown on succinate-ammonium chloride did not possess the transport system, but it could be induced in such cells by methylamine in 20 h. Cells grown with methylamine as a sole nitrogen, but not carbon, source transported methylamine at a reduced rate.
Construction and characterization of Escherichia coli polA-lacZ gene fusions
Authors: Ward D F , Murray N E
Published: January 2020

The promoter of the polA gene of Escherichia coli K-12 was fused to the lacZ gene by selecting deletions within a lambda lacZ polA transducing phage. Four fusions, deleting varying amounts of the polA gene, were characterized. The polA promoter was found to be approximately 3% as active as the fully induced lac promoter. This figure is compatible with the normal intracellular level of deoxyribonucleic acid polymerase I. No evidence was found for outogenous regulation of transcription from the polA promoter. Expression from this promoter was influenced by neither recA nor mitomycin C, but uvrD and uvrE mutations reduced expression slightly.
Bacteriophage T4D Receptor and the
Escherichia coli
Cell Wall Structure: Binding of Endotoxin-Like Particles to the Cell Wall
Authors: Zorzopulos Jorge , DeLong Sara , Chapman Virginia , Kozloff Lloyd M.
Published: January 2020

A variety of degradative treatments have been used to investigate the nature of the structure and components of the cell walls of
Escherichia coli
B. The binding and localization of the endotoxin-like particles found on the cell walls were of special interest because some of them are associated with the site where the inner tail tube of bacteriophage T4D penetrates the cell wall. Modified cell walls were obtained by heating a suspension of bacterial cells originally in 0.1 M phosphate, pH 7.0, after the addition of 12.5 M NaOH to a final concentration of 0.25 M. With regard to the endotoxin-like particles, it was found that: (i) at least part of them still remained bound to the modified cell wall after the alkali treatment; (ii) the subsequent incubation of alkali-treated cell walls with lysozyme destroyed the bacterial form and released a complex of endotoxin-like particles together with a fibrous material; (iii) on the other hand, treatment with 45% phenol at 70°C removed the endotoxin-like particles from the surface of the alkali-treated cell walls, but most of the fibrous material was left on the cell wall; and (iv) incubation of alkali-treated cell walls with 5 mM ethylenediaminetetraacetic acid at 20°C also removed the endotoxin-like particles, but did not disrupt the rodlike bacterial form. However, if the ethylenediaminetetraacetic acid treatment was performed at 55°C, the bacterium-like form was destroyed. These differential sensitivities to ethylenediaminetetraacetic acid suggested that loosely bound divalent metal ions normally hold these endotoxin-like particles on the cell wall surface, but that probably more tightly bound metal ions are involved in the determination of cell shape. Analysis of the protein components of the alkalitreated cell walls showed that only one protein was present in significant amounts, and this protein had an electrophoretic mobility similar to that of the Braun lipoprotein. This protein was released from the alkali-treated cell walls upon heating with 2% sodium dodecyl sulfate at 100°C. Phospholipids were also absent from this structure. The distribution of the remaining cell wall components on the alkali-treated cell walls is discussed.
Cross-linking analysis of Neisseria gonorrhoeae outer membrane proteins
Authors: Leith D K , Morse S A
Published: January 2020

The arrangement of proteins in the outer membrane of Neisseria gonorrhoeae was investigated through the use of cleavable chemical cross-linking reagents and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cross-linking of isolated outer membranes yielded dimers and trimers of the major outer membrane protein. In addition, data were obtained suggesting that a stable interaction exists between the major protein I and protein II, the second most prevalent protein in the gonococcal outer membrane.