bluuebunny/crossref_metadata_2025 huggingface dataset:
Degradation of 4-chlorophenylacetic acid by a Pseudomonas species
Authors: Klages U , Markus A , Lingens F
Published: January 2020

Pseudomonas sp. strain CBS3 was able to utilize 4-chlorophenylacetic acid as the sole source of carbon and energy. When this strain was grown with 4-chlorophenylacetic acid, homoprotocatechuic acid was found to be an intermediate which was further metabolized by the meta-cleavage pathway. Furthermore, three isomers of chlorohydroxyphenylacetic acid, two of them identified as 3-chloro-4-hydroxyphenylacetic acid and 4-chloro-3-hydroxyphenylacetic acid, were isolated from the culture medium. 4-Hydroxyphenylacetic acid was catabolized in a different manner by the glutathione-dependent homogentisate pathway. Degradation enzymes of both of these pathways were inducible.
R-factor cointegrate formation in Salmonella typhimurium bacteriophage type 201 strains
Authors: Helmuth R , Stephan R , Bulling E , van Leeuwen W J , van Embden J D , Guinée P A , Portnoy D , Falkow S
Published: January 2020

The genetic and molecular properties of the plasmids in Salmonella typhimurium phase type 201 isolated are described. Such strains are resistant to streptomycin, tetracycline, chloramphenicol, ampicillin, kanamycin, and several other antimicrobial drugs, and are highly pathogenic for calves. These strains have been encountered with increasing frequency since 1972 in West Germany and The Netherlands. We show that isolates of this phage type constitute a very homogeneous group with regard to their extrachromosomal elements. These bacteria carry three small plasmids: pRQ3, a 4.2-megadalton (Md) colicinogenic plasmid; pRQ4, 3.4-Md plasmid that interferes with the propagation of phages; and pRQ5, a 3.2-Md cryptic plasmid. Tetracycline resistance resides on a conjugative 120-MD plasmid pRQ1, belonging to the incompatibility class H2. Other antibiotic resistance determinants are encoded by a nonconjugative 108-Md plasmid pRQ2. Transfer of multiple-antibiotic resistance to appropriate recipient strains was associated with the appearance of a 230-Md plasmid, pRQ6. It appears that pRQ6 is a stable cointegrate of pRQ1 and pRQ2. This cointegrate plasmid was transferable with the same efficiency as pRQ1. Other conjugative plasmids could mobilize pRQ2, but stable cointegrates were not detected in the transconjugants. Phase type 201 strains carry a prophage, and we show that phage pattern 201 reflects the interference with propagation of typing phages effected by this prophage and plasmid pRQ4 in strains of phage type 201.
Metabolism of resorcinylic compounds by bacteria: new pathway for resorcinol catabolism in Azotobacter vinelandii
Authors: Groseclose E E , Ribbons D W
Published: January 2020

We present evidence to document a third pathway for the microbial catabolism of resorcinol. Resorcinol is converted to pyrogallol by resorcinol-grown cells of Azotobacter vinelandii. Pyrogallol is the substrate for one of two ring cleavage enzymes induced by growth with resorcinol. Oxalocrotonate, CO2, pyruvate, and acetaldehyde have been identified as products of pyrogallol oxidation catalyzed by extracts of resorcinol-grown cells. The enzymes pyrogallol 1,2-dioxygenase, oxalocrotonate tautomerase (isomerase), oxalocrotonate decarboxylase, and vinylpyruvate hydratase are present in extracts from resorcinol-grown cells but not in succinate-grown cells.
Metabolism of sulfur compounds by whole filaments and heterocysts of Anabaena variabilis
Authors: Giddings T H , Wolk C P , Shomer-Ilan A
Published: January 2020

Filaments of the heterocyst-forming cyanobacterium Anabaena variabilis reproduced 35SO4(2)-, incorporating 35S into cysteine, methionine, glutathione, sulfolipid, and several unidentified metabolites. The majority of the incorporated label accumulated in reduced glutathione. Heterocysts isolated from labeled filaments contained the same major labeled products. Isolated, metabolically active heterocysts were unable to reduce 35SO4(2)-, but were able to incorporate 35S2- into cysteine and glutathione. The results suggest that the initial activation of SO4(2)- occurs in vegetative cells and that some reduced forms, possibly including S20, are translocated into heterocysts.
Failure of
Escherichia coli
to Alter Its Fatty Acid Composition in Response to Cholesterol-Induced Changes in Membrane Fluidity
Authors: Eaton L. C. , Erdos G. W. , Vreeland N. L. , Ingram L. O.
Published: January 2020

Substantial amounts of exogenously supplied cholesterol were incorporated into the membranes of
Escherichia coli
during growth and caused a large decrease in membrane fluidity. Although no compensatory changes in fatty acid composition were observed, the incorporation of cholesterol did not affect the rate of growth of
E. coli
or interfere with the changes in fatty acid composition which normally occur during growth at different temperatures.
Isolation and characterization of spontaneously occurring TOL plasmid mutants of Pseudomonas putida HS1
Authors: Kunz D A , Chapman P J
Published: January 2020

A strain of Pseudomonas (P. putida HS1) was found to resemble P. putida (arvilla) mt-2 in its ability to degrade toluene, m- and p-xylene, 1,2,4-trimethylbenzene (pseudocumene), and 3-ethyltoluene via oxidation of a methyl substituent and reactions of the meta-fission pathway. The ability to degrade these substrates by P. putida HS1 (PpC1) was shown to be encoded by a TOL (pDK1) plasmid as evidenced by: (i) spontaneous loss of the TOL-related phenotype after growth with benzoate, (ii) transfer of the TOL character from the wild type into cured recipients by conjugation, and (iii) isolation of a plasmid of identical molecular weight (120 X 10(6)) from both the wild type and an exconjugant obtained by mating wild type with a putative cured recipient. In addition to the isolation of apparent cured strains having lost the entire TOL-related phenotype, two additional mutant classes were observed after growth on benzoate. One class, represented by PpCT1, was unable to utilize the alkyl-substituted aromatic compounds but retained the ability to grow with toluene and benzyl alcohol. Analysis of PpCT1 revealed that it was unable to synthesize the TOL-encoded toluate oxidase and enzymes of the meta pathway but retained the ability to elaborate activities for toluene hydroxylase, benzyl alcohol, and benzaldehyde dehydrogenase, thereby mediating initial oxidation of toluene to benzoate, which was then further metabolized via enzymes of the chromosomally encoded ortho-fission pathway. A second class of mutants had lost the ability to utilize the hydrocarbons but could still grow with m-toluate but not p-toluate, 3,4-dimethylbenzoate, or 3-ethylbenzoate, intermediates in the oxidation of the corresponding hydrocarbons. Our such mutant, PpCM1, could no longer synthesize enzymes required for initial oxidation of the hydrocarbons, but was able to produce the toluate oxidase and enzymes of the meta pathway, thereby facilitating degradation of m-toluate. Neither PpCT1, PpCM1, nor a putative cured strain, PpCC1, reverted at detectable frequencies (less than 10(-9). Analysis of each strain for plasmid deoxyribonucleic acid revealed the presence of a single plasmid in each strain with the following molecular weights: PpCM1, 100 X 10(6) (pDKM1); PpCT1, 80 X 10(6) (pDKT1); PpCC1 20 X 10(6) (pDKC1). The results suggest that the TOL (pDK1) plasmid has undergone deletions giving rise to smaller replicons which either encode for only a fraction of the wild-type catabolic functions (pDKM1, pDKT1) or have lost all catabolic activities (pDKC1).
Localization of phoE, the structural gene for outer membrane protein e in Escherichia coli K-12
Authors: Tommassen J , Lugtenberg B
Published: January 2020

To localize the structural gene for outer membrane protein e, mutants resistant to the protein e-specific phage TC45 were isolated and characterized. Three classes of TC45-resistant strains were found: (i) mutants in phoB, a regulatory gene for protein e, (ii) mutants with an altered lipopolysaccharide, and (iii) mutants unaltered in the regulation of the pho regulon and producing an apparently normal lipopolysaccharide. Mutations in the latter class of mutants are probably altered in the structural gene for protein e and are cotransducible with proA,B at min 6 on the chromosomal map. As class (iii) includes mutants with an electrophoretically altered protein e, in both an nmpA and an nmpB background, we conclude that the structural gene for protein e, designated as phoE, is localized at min 6 on the chromosomal map, the gene order being phoE proA argF.
Isolation of a plasmid responsible for caseinase activity in Clostridium perfringens ATCC 3626B
Authors: Blaschek H P , Solberg M
Published: January 2020

Clostridium perfringens strain ATCC 3626B was cured of caseinase activity at a high frequency after treatment with acriflavine dye (2.5%) or elevated temperature growth (9.1%). Caseinase-negative isolates retained the larger (9.4 megadaltons) pHB102 cryptic plasmid, but were missing the smaller (2.1 megadaltons) pHB101 plasmid present in the caseinase-positive wild-type strain. Dye-buoyant density-gradient centrifugation at 4 or 15 degrees C revealed that the pHB101 and pHB102 plasmids are temperature labile and easily converted into the nicked non-supercoiled or linear state.
Mode of elongation of the glycerol phosphate polymer of membrane lipoteichoic acid of Streptococcus faecium ATCC 9790
Authors: Cabacungan E , Pieringer R A
Published: January 2020

Specific degradation of membrane lipoteichoic acid of Streptococcus faecium ATCC 9790 by a phosphodiesterase from Aspergillus niger and by periodate oxidation has demonstrated that the enzymatic synthesis of the glycerol phosphate polymer of the molecule occurs by an external elongation system. Evidence of this type of mechanism was obtained with lipoteichoic acid synthesized in vivo or in vitro by differential radioisotope labeling techniques. The glycerol phosphate repeating units were transferred from phosphatidylglycerol and became linked through a phosphodiester bond to the glycerol phosphate unit of the chain farthest from or most external to the lipid end of the polymer.
Regulation of L-asparaginase in a Chlamydomonas species in response to ambient concentrations of combined nitrogen
Authors: Paul J H , Cooksey K E
Published: January 2020

Cellular levels of an L-asparaginase in a Chlamydomonas species were found to be greater in nitrogen-limited batch cultures than in batch cultures grown in ample nitrogen. Cells grown in high nitrogen medium (5 mM NH4Cl) and suspended in nitrogen-free medium showed a 2- to 3.5-fold increase in activity after 24 to 48 h. This increase in activity was inhibited by cycloheximide and by the addition of high levels of combined nitrogen (5 mM NH4Cl, NaNO3, or L-asparagine), suggesting repression by ambient nitrogen levels as the mode of regulation of this enzyme. Derepressed L-asparaginase activity did not disappear in the presence of high concentrations of medium nitrogen, indicating the absence of an asparaginase-degrading system. Derepression of asparaginase by this organism was light dependent and inhibited by 3-(3',4'-dichlorophenyl)-1,1-dimethylurea suggesting a requirement for photosynthetic energy.
Physical map of the Salmonella typhimurium histidine transport operon: correlation with the genetic map
Authors: Ardeshir F , Higgins C F , Ames G F
Published: January 2020

A detailed restriction map of a 12.4-kilobase EcoRI fragment of Salmonella typhimurium deoxyribonucleic acid (DNA) containing the entire histidine transport operon and the argT gene is presented. Subclones of specific regions of the transport operon of S. typhimurium were constructed in plasmid vectors. An accurate correlation between the restriction map and the location of genetically defined deletions was obtained by hybridizing restriction digests of chromosomal DNA from strains carrying each deletion with cloned transport operon DNA as a probe. These data were used to position the histidine transport genes on the cloned 12.4-kilobase fragment of DNA.
Carboxypeptidase S- and Carboxypeptidase Y-Deficient Mutants of
Saccharomyces cerevisiae
Authors: Wolf Dieter H. , Ehmann Claudia
Published: January 2020

A new carboxypeptidase (carboxypeptidase S) was found in a
Saccharomyces cerevisiae
strain lacking carboxypeptidase Y (D. H. Wolf and U. Weiser, Eur. J. Biochem.
73:
553-556, 1977). Mutants devoid of carboxypeptidase S activity were isolated from a mutant strain that was also deficient in carboxypeptidase Y. Four mutants were analyzed in detail and fell into one complementation group. The defect segregated 2:2 in meiotic tetrads. Gene dosage experiments indicated that the mutation might reside in the structural gene of carboxypeptidase S. The absence of both enzymes, carboxypeptidases Y and S, did not affect mitotic growth. Ascopore formation was only slightly affected by the absence of both carboxypeptidases. Protein degradation under conditions of nutrient deprivation and under sporulation conditions showed no obvious alteration in the absence of carboxypeptidases Y and S. When a proteinase B mutation, which led to the absence of proteinase B activity and resulted in the partial reduction of sporulation, was introduced into a mutant lacking both carboxypeptidases, the ability of diploid cells to sporulate was nearly completely lost. Mutants lacking both carboxypeptidases were unable to grow on the dipeptide benzyloxycarbonylglycyl-
l
-leucine as a sole nitrogen source, which indicates an additional function for carboxypeptidases Y and S in supplying nutrients from exogenous peptides. Catabolite inactivation of fructose-1,6-bisphosphatase, cytoplasmic malate dehydrogenase, and phosphoenolpyruvate carboxykinase and inactivation of nicotin-amide adenine dinucleotide phosphate-dependent, glutamate dehydrogenase, events which have been proposed to involve proteolysis in vivo, were not dependent on the presence of carboxypeptidase Y and S. In a mutant lacking both carboxypeptidases, four new proteolytic enzymes with carboxypeptidase activity were detected.
Effects of beta-lactam antibiotics on peptidoglycan synthesis in growing Neisseria gonorrhoeae, including changes in the degree of O-acetylation
Authors: Blundell J K , Perkins H R
Published: January 2020

Low concentrations of beta-lactam antibiotics caused an increased uptake of radioactive glucosamine into the sodium dodecyl sulfate-insoluble peptidoglycan of growing Neisseria gonorrhoeae. There was no appreciable change in the (small) amount of sodium dodecyl sulfate-soluble polymer present in the cultures. The sodium dodecyl sulfate-insoluble product in control cells was only partially dissolved by egg-white lysozyme (about 40%), but could all be released by the Chalaropsis B muramidase. In cells exposed to beta-lactams the proportion of labeled peptidoglycan susceptible to lysozyme increased to 60%. Examination of the Chalaropsis B digests by thin-layer chromatography showed that they contained disaccharide-peptide monomers with and without O-acetylation and bis-disaccharide-peptide dimers with one or two O-acetyl groups, or with none. beta-Lactam antibiotics caused a decrease in the degree of O-acetylation but did not greatly affect the amount of peptidoglycan cross-linking. They also had the effect of enlarging the bacteria and conserving and thickening the septa that could be observed in thin sections under the electron microscope. The relationship between these results and the effects of beta-lactams on in vitro synthesis of peptidoglycan by ether-treated N. gonorrhoeae is discussed.
A Pseudomonas aeruginosa mutant non-derepressible for orthophosphate-regulated proteins
Authors: Gray G L , Berka R M , Vasil M L
Published: January 2020

Using a rapid screening assay based on the hydrolysis of p-nitrophenylphosphorylcholine, we isolated several mutants of Pseudomonas aeruginosa deficient in the production of phospholipase C. One, designated strain A50N, was also markedly deficient in the synthesis of alkaline phosphatase and several unidentified extracellular proteins. Because strain A50N produces these proteins under conditions of derepression at levels equal to those produced by the parental strain PAO1 grown in medium containing excess phosphate, it appears to have a mutation in a genetic element involved in the derepression of phosphate-repressible proteins.
Lipid Intermediate in the Synthesis of the Linkage Unit That Joins Teichoic Acid to Peptidoglycan inBacillus subtilis
Authors: Hancock Ian C. , Wiseman Gordon , Baddiley James
Published: January 2020

Membranes fromBacillus subtilisW23 synthesized a lipid precursor of the linkage unit that attaches teichoic acid to the cell wall. It contained glycerophosphoryl-N-acetylglucosamine, linked through an acid-labile bond to a lipid.
The tsx protein of Escherichia coli can act as a pore for amino acids
Authors: Heuzenroeder M W , Reeves P
Published: January 2020

The tsx protein is known to be a specific diffusion pathway for nucleosides. The ability of this protein to facilitate the transport of molecules other than nucleosides was examined in strains lacking detectable amounts of porin (ompB mutants). The tsx protein was shown to promote serine, glycine, and phenylalanine transport and to have no effect on either glucose or arginine transport.
Escherichia coli mutants in which transcription is dependent on recA function
Authors: Froehlich B , Epstein W
Published: January 2020

A gene essential for viability in recA mutants of Escherichia coli K-12 was identified. This gene, rdg for recA-dependent growth, is near 16 min on the E. coli chromosome between phr and gltA and is 90% cotransduced with gltA. In a strain with an rdg deletion and the temperature-sensitive recA allele, recA200, growth stopped within 7 min after cells were shifted to the nonpermissive temperature (42 degrees C). The cells remained viable for many hours at 42 degrees C. The defect at the nonpermissive temperature is in ribonucleic acid synthesis, which was completely shut off within 20 min after the temperature shift. Protein synthesis was also shut off, but deoxyribonucleic acid synthesis continued for at least 2 h after the shift. The rdg mutation alone had no apparent effect on growth, deoxyribonucleic acid repair, or recombination.
Adenosine 5′-Triphosphate-Yielding Pathways of Branched-Chain Amino Acid Fermentation by a Marine Spirochete
Authors: Harwood C. S. , Canale-Parola E.
Published: January 2020

The metabolic pathways utilized by an obligately anaerobic marine spirochete (strain MA-2) to ferment branched-chain amino acids were studied. The spirochete catabolized
l
-leucine to isovaleric acid,
l
-isoleucine to 2-methylbutyric acid, and
l
-valine to isobutyric acid, with accompanying CO
2
production in each fermentation. Cell extracts of spirochete MA-2 converted
l
-leucine,
l
-isoleucine, and
l
-valine to 2-ketoisocaproic, 2-keto-3-methylvaleric, and 2-ketoisovaleric acids, respectively, through mediation of 2-ketoglutarate-dependent aminotransferase activities. The branched-chain keto acids were decarboxylated and oxidized to form isovaleryl coenzyme A, 2-methylbutyryl coenzyme A, and isobutyryl coenzyme A, respectively, in the presence of sulfhydryl coenzyme A and benzyl viologen. The acyl coenzyme A's were converted to acyl phosphates by phosphate branched-chain acyltransferase enzymatic activities. Branched-chain fatty acid kinase activities catalyzed formation of isovaleric, 2-methylbutyric, and isobutyric acids from isovaleryl phosphate, 2-methylbutyryl phosphate, and isobutyryl phosphate, respectively. Adenosine 5′-triphosphate was formed during conversion of branched-chain acyl phosphates to branched-chain fatty acids. The results indicate that conversion of
l
-leucine,
l
-isoleucine, and
l
-valine to branched-chain fatty acids by spirochete MA-2 results in adenosine 5′-triphosphate generation. The metabolic pathways utilized for this conversion involve amino acid amino-transferase, 2-keto acid oxidoreductase, phosphate acyltransferase, and fatty acid kinase activities.
Exogenous induction of the Escherichia coli hexose phosphate transport system defined by uhp-lac operon fusions
Authors: Shattuck-Eidens D M , Kadner R J
Published: January 2020

The uhp-coded hexose phosphate transport system of Escherichia coli is normally induced by the presence of extracellular glucose-6-phosphate (G6P), whereas internally generated G6P does not provide a regulatory signal. Strains carrying uhp-lac operon fusions in which lac operon expression is under the control of the uhpT promoter were isolated. The direction of transcription of the uhp T gene was found to be counterclockwise on the E. coli chromosome map. The effects of added sugar phosphates on induction of beta-galactosidase and G6P uptake activities were compared in two fusion-carrying strains differing only in the presence of functional Uhp+ activity. Induction of uhp expression by G6P was equally effective in the two strains; accumulation of G6P diminished its ability to serve as an inducer. Mannose-6-phosphate was an effective competitive inhibitor of G6P uptake, but did not inhibit induction by G6P of uhp expression. No sugar phosphates were found that inhibited induction by G6P. Inorganic phosphate competitively inhibited induction by G6P whether G6P transport activity was present or not. Thus, the transport activity is not involved in the regulation of its synthesis, and these results strongly support the view that the uhp regulatory system senses only the external environment.
Proline transport in Saccharomyces cerevisiae
Authors: Lasko P F , Brandriss M C
Published: January 2020

The yeast Saccharomyces cerevisiae is capable of utilizing proline as the sole source of nitrogen. Mutants of S. cerevisiae with defective proline transport were isolated by selecting for resistance to either of the toxic proline analogs L-azetidine-2-carboxylate or 3,4-dehydro-DL-proline. Strains carrying the put4 mutation are defective in the high-affinity proline transport system. These mutants could still grow when given high concentrations of proline, due to the operation of low-affinity systems whose existence as confirmed by kinetic studies. Both systems were repressed by ammonium ions, and either was induce by proline. Low-affinity transport was inhibited by histidine, so put4 mutants were unable to grow on a medium containing high concentrations of proline to which histidine has been added.
Genetic transformation in Staphylococcus aureus: demonstration of a competence-conferring factor of bacteriophage origin in bacteriophage 80 alpha lysates
Authors: Thompson N E , Pattee P A
Published: January 2020

A virion component that is responsible for conferring competence to Staphylococcus aureus was demonstrated in lysates of bacteriophage 80 alpha, a serological group B phage. Isolated particles of 80 alpha could not be shown to confer significant levels of competence. The phage component had a density of about 1.3 g/cm3, was inactivated by pronase, and was inhibited by antiserum prepared against isolated infectious particles of a serological group B phage. Centrifugation through a Ficoll gradient resulted in separation of competence-conferring activity and plaque-forming units. It is concluded that this proteinaceous subvirion component constitutes a bona fide competence factor of bacteriophage origin.
Gliding motility in Aphanothece halophytica: analysis of wall proteins in mot mutants
Authors: Simon R D
Published: January 2020

The unicellular cyanobacterium Aphanothece halophytica (PCC 7418) is motile, and spontaneous nonmotile (mot) mutants accumulate when the organism is subcultured. Analysis of mot mutants suggests that a glycoprotein in the cell wall is involved in the motility mechanism. Proteins from the wall fraction of the wild type and five mot clones were analyzed by gradient sodium dodecyl sulfate-acrylamide gel electrophoresis. Four clones were similar to the wild type, and one clone, mot-3, was missing a high-molecular-weight protein (approximately 200,000) and had at least one new polypeptide (160,000). The high-molecular-weight protein stained with periodic acid-Schiff reagent, suggesting that it was a glycoprotein. The absence of the protein in mot-3 did not affect the mechanical strength of the wall, since both mot-3 and wild-type cells were broken at the same rate by controlled cavitation. Several other cyanobacteria were also screened for the presence of glycoproteins. All motile strains have such proteins, although none had an apparent molecular weight as high as that in Aphanothece sp. Some motile strains, such as Oscillatoria limnetica and Phormidium sp., showed very large amounts of glycoproteins; whereas some nonmotile strains, such as Synechococcus sp. (UTEX 625) and Microcystis sp. (PCC 7820), showed no high-molecular-weight glycoproteins.
Properties of crystalline L-ornithine: alpha-ketoglutarate delta-aminotransferase from Bacillus sphaericus
Authors: Yasuda M , Tanizawa K , Misono H , Toyama S , Soda K
Published: January 2020

The distribution of bacterial L-ornithine: alpha-ketoglutarate delta-aminotransferase (L-ornithine:2-oxo-acid aminotransferase [EC 2.6.1.13]) was investigated, and Bacillus sphaericus (IFO 3525) was found to have the highest activity of the enzyme, which was inducibly formed by addition of L-ornithine or L-arginine to the medium. L-Ornithine:alpha-ketoglutarate delta-aminotransferase, purified to homogeneity and crystallized from B. sphaericus, had a molecular weight of about 80,000 and consisted of two subunits identical in molecular weight (41,000) and in amino-terminal residue (threonine). The enzyme exhibited absorption maxima at 278,343, and 425 nm and contained 1 mol of pyridoxal 5'-phosphate per mol of enzyme. The formyl group of pyridoxal 5'-phosphate was bound through an aldimine linkage to the epsilon-amino group of a lysine residue of the protein. The enzyme-bound pyridoxal 5'-phosphate, absorbing at 425 nm, was released by incubation with phenylhydrazine to yield the catalytically inactive form. The inactive enzyme, which was reactivated by addition of pyridoxal 5'-phosphate, still had a 343-nm peak and contained 1 mol of a vitamin B6 compound. The holoenzyme showed positive circular dichroic bands at 340 and 425 nm, whereas the inactive form had no band at 425 nm. The enzyme was highly specific for L-ornithine and alpha-ketoglutarate and catalyzed delta-transamination between them to produce L-glutamate and L-glutamate-gamma-semialdehyde, which as spontaneously converted to delta 1-pyrroline-5-carboxylate. The enzyme activity was significantly affected by nonsubstrate amino acids, amines, and carbonyl reagents.
Increased outer membrane resistance to ethylenediaminetetraacetate and cations in novel lipid A mutants
Authors: Vaara M
Published: January 2020

Polymyxin-resistant pmrA mutants of Salmonella typhimurium differed from their parents in that they were resistant to tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-lysozyme, tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-deoxycholate, and tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate-bacitracin. Tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetate released about 50% less lipopolysaccharide from the pmrA strains than from the parental strains when the bacteria were grown in L-broth containing 2 mM Ca2+. Protamine, polylysine, octapeptin, benzalkonium chloride, cold NaCl, cold MgCl2, or cold tris(hydroxymethyl)aminomethane hydrochloride (pH 7.2) caused no leakage or markedly less leakage of periplasmic beta-lactamase from a pmrA mutant than from its parent strain. pmrA mutants were more resistant than their parent strains to protamine and polylysine but not to octapeptin or benzalkonium chloride, as measured by the ability of these agents to kill the bacteria or to sensitize them to deoxycholate-induced lysis. The pmrA strains did not differ from their parent strains in sensitivity to several antibiotics, in porin function (as measured by cephaloridine diffusion across the outer membrane), or in outer membrane-associated phospholipase A activity.
Differential methicillin susceptibilities of peptidoglycan syntheses in methicillin-resistant Staphylococcus aureus
Authors: Smith P F , Wilkinson B J
Published: January 2020

The mechanism of staphylococcal resistance to methicillin is unknown. Peptidoglycan synthesis was studied in a methicillin-resistant and a derived methicillin-sensitive Staphylococcus aureus strain. Although the methicillin minimum inhibitory concentration for growth of the methicillin-resistant strain was 1,600 micrograms/ml, peptidoglycan synthesis by the organism incubated in a wall synthesis solution was inhibited about 90% by 5 micrograms of methicillin per ml. In contrast, high concentrations of methicillin added to actively growing cultures of the methicillin-resistant strain had little effect on growth or peptidoglycan synthesis. Peptidoglycan synthesis in chloramphenicol-treated cultures was more susceptible to methicillin than it was in actively growing cultures of the methicillin-resistant strain. It is proposed that in this strain cell wall thickening peptidoglycan synthesis which predominates in cell wall synthesis solution and chloramphenicol-treated cultures is methicillin sensitive, whereas peptidoglycan synthesis involved in cell division, primarily in the region of the septum, which predominates in actively growing cultures is methicillin resistant. Both cell wall thickening and septal peptidoglycan syntheses are methicillin sensitive in the methicillin-sensitive strain.
Development-specific protein S of Myxococcus xanthus: purification and characterization
Authors: Inouye S , Harada W , Zusman D , Inouye M
Published: January 2020

Protein S, a development-specific protein of Myxococcus xanthus, was purified from the cells of a late stage of development and crystallized. Its circular dichroism spectra indicated that protein S had a high content of beta-structure in both the presence and absence of calcium ion, which is required for self-assembly of protein S on the myxospore surface. Its amino and carboxyl terminal sequences were determined to be alanine-aspartic acid-isoleucine-glycine-valine-alanine-methionine-asparagine-asparagine-aspartic acid-threonine-serine-serine and isoleucine-arginine (isoleucine, serine), respectively. When protein S (molecular weight, 23,000) was digested with trypsin, a trypsin-resistant core of 10,000 molecular weight was obtained. The core peptide was purified, and its amino acid composition was compared with that of protein S. The core peptide was capable of self-assembly on the spore surface in the presence of calcium ion and competed with protein S for binding on the spore surface. The ratio of affinity to the spore surface for protein S to that for the core peptide was 1.55.
Effect of nalidixic acid and novobiocin on pBR322 genetic expression in Escherichia coli minicells
Authors: Gómez-Eichelmann M C
Published: January 2020

The effects of two deoxyribonucleic acid (DNA) gyrase inhibitors, nalidixic acid and novobiocin, on the gene expression of plasmid pBR322 in Escherichia coli minicells were studied. Quantitative estimates of the synthesis of pBR322-coded polypeptides in novobiocin-treated minicells showed that the synthesis of a polypeptide of molecular weight of 34,000 (the tetracycline resistance protein) was reduced to 11 to 20% of control levels, whereas the amount of a polypeptide of 30,500 (the beta-lactamase precursor) was increased to as much as 200%. Nalidixic acid affected the synthesis of the tetracycline resistance protein similarly to novobiocin, although to a lesser extent. The effects of nalidixic acid were not observed in a nalidixic-resistant mutant; those induced by novobiocin were only partially suppressed in a novobiocin-resistant mutant. The synthesis of one of the inducible tetracycline-resistant proteins (34,000) coded by plasmid pSC101 was also reduced in nalidixic acid- and novobiocin-treated minicells. These results suggest that the gyrase inhibitors modified the interaction of ribonucleic acid polymerase with some promoters, either by decreasing the supercoiling density of plasmid DNA or by altering the association constant of the gyrase to specific DNA sites.
In vivo and in vitro functional alterations of the bacteriophage lambda receptor in lamB missense mutants of Escherichia coli K-12
Authors: Braun-Breton C , Hofnung M
Published: January 2020

lamB is the structural gene for the bacteriophage lambda receptor in Escherichia coli K-12. In vivo and in vitro studies of the lambda receptor from lamB missence mutants selected as resistant to phage lambda h+ showed the following. (i) Resistance was not due to a change in the amount of lambda receptor protein present in the outer membrane but rather to a change in activity. All of the mutants were still sensitive to phage lambda hh*, a two-step host range mutant of phage lambda h+. Some (10/16) were still sensitive to phage lambda h, a one-step host range mutant. (ii) Resistance occurred either by a loss of binding ability or by a block in a later irreversible step. Among the 16 mutations, 14 affected binding of lambda h+. Two (lamB106 and lamB110) affected inactivation but not binding; they represented the first genetic evidence for a role of the lambda receptor in more than one step of phage inactivation. Similarly, among the six mutations yielding resistance to lambda h, five affected binding and one (lamB109) did not. (iii) The pattern of interactions between the mutated receptors and lambda h+ and its host range mutants were very similar, although not identical, in vivo and in vitro. Defects were usually more visible in vitro than in vivo, the only exception being lamB109. (iv) The ability to use dextrins as a carbon source was not appreciably affected in the mutants. Possible working models and the relations between phage infection and dextrins transport were briefly discussed.
Thiolation and 2-methylthio- modification of Bacillus subtilis transfer ribonucleic acids
Authors: Vold B S , Longmire M E , Keith D E
Published: January 2020

Six thionucleosides found in Bacillus subtilis transfer ribonucleic acids were investigated: N6-(delta 2-isopentenyl)-2-methylthioadenosine, 5-carboxymethylaminomethyl-2-thiouridine, 4-thiouridine, 2-methylthioadenosine, N-[(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl]threonine, and one unknown (X1). The presence of N-[(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl]threonine was demonstrated based on the affinity of the transfer ribonucleic acid containing it for an immunoadsorbent made with the antibody directed toward N-[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]-L-threonine. The existance of N-[(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl]threonine in two species of lysine transfer ribonucleic acids was also confirmed by high-resolution mass spectrometry. Four of these thionucleosides--N6-(delta 2-isopenenyl)-2-methylthioadenosine, 2-methylthioadenosine, 5-carboxymethylaminomethyl-2-thiouridine, and the unknown designated X1--occurred only in specific areas in the elution profile of an RPC-5 column and probably affect the chromatographic properties of the transfer ribonucleic acids containing them. In contrast with Escherichia coli, where 4-thiouridine is the most frequent type of sulfur-containing modification, approximately one-third of the sulfur groups in B. subtilis transfer ribonucleic acid are present as thiomethyl groups on the 2 position of an adenosine or modified adenosine residue.
Curing of the killer deoxyribonucleic acid plasmids of Kluyveromyces lactis
Authors: Niwa O , Sakaguchi K , Gunge N
Published: January 2020

Ultraviolet irradiation gave rise to frequent curing of killer plasmids pGKl1 and pGK12 of Kluyveromyces lactis. Almost all of the nonkillers obtained lost both plasmids, but one of them lost only pGKl1. The disappearance of pGKl1 was accompanied by the simultaneous loss of the killer activity and of the resistance to the killer factor. A new plasmid, pGKl1S, was obtained, which arose from a deletion in the central region of pGKl1. Genetic analysis suggested that pGKl1S has the killer gene lost by the deletion and the resistance gene intact and that pGKl1S shares the same replication control with pGKl1.
Beta-lactamase-specifying plasmids isolated from Neisseria gonorrhoeae have retained an intact right part of a Tn3-like transposon
Authors: Fayet O , Froment Y , Piffaretti J C
Published: January 2020

In three beta-lactamase-producing strains of Neisseria gonorrhoeae, ampicillin resistance is due to the presence of a 7.4-kilobase plasmid. Heteroduplex analysis has shown that the R plasmid contains a 1.6-kilobase segment homologous to the right part (the region coding for the beta-lactamase) of the Tn3-like transposon Tn2301 the 1.6-kilobase DNA segment is not transposable, but it can give rise to a functional transposon, when linked to the left part of TN2301. This provides strong evidence that the R plasmids of N. gonorrhoeae are deletion derivatives of a plasmid that contained an entire TN3-like transposon.
Cloning and expression in Escherichia coli K-12 of the genes for major outer membrane protein OmpA from Shigella dysenteriae, Enterobacter aerogenes, and Serratia marcescens
Authors: Cole S T , Sonntag I , Henning U
Published: January 2020

The outer membranes of many gram-negative bacteria contain a major heat-modifiable protein which shows serological cross-reactivity with the OmpA protein of Escherichia coli K-12. Using the cloned gene for the E. coli K12 protein as a DNA-DNA hybridization probe, we were able to identify the corresponding genes from Shigella dysenteriae. Enterobacter aerogenes, and Serratia marcescens. These were cloned in a phage lambda vector, and their expression in E. coli K-12 was studied. All three OmpA proteins were fully produced and correctly exported to the outer membrane. In several cases, complete or partial restoration of known function of the E. coli K-12 protein was observed.
Genetic regulation of the glyoxylate shunt in Escherichia coli K-12
Authors: Maloy S R , Nunn W D
Published: January 2020

The expression of the glyoxylate shunt enzymes is required for growth of Escherichia coli on acetate or fatty acids as a sole carbon source. The genes for the two unique enzymes of the glyoxylate shunt, aceA and aceB, are located at 90 min on the E. coli K-12 genetic map. Polar mutations in the aceB gene eliminate aceA gene function, suggesting that these genes constitute an operon and the direction of transcription is from aceB to aceA. Mu d (Ap lac) fusions with the aceA gene have been constructed to study the regulation of the ace operon. Expression of the ace operon is under the transcriptional control of two genes: the iclR gene, which maps near the ace operon, and the fadR gene, which maps at 25 min, and is also involved in the regulation of the fatty acid degradation (fad) regulon. Merodiploid studies demonstrated that both the iclR and fadR genes regulate the glyoxylate shunt in a trans-dominant manner.
Spontaneous protoplast formation in Methanobacterium bryantii
Authors: Jarrell K F , Colvin J R , Sprott G D
Published: January 2020

Methanobacterium bryantii was found to undergo rapid lysis when grown in a prereduced chemically defined medium under H2-CO2 (4:1, vol/vol). The addition of 20 mM MgCl2 to the medium gave, rather than rapid lysis, a gradual formation of phase-dark spherical bodies which in thin section appeared as true protoplasts. In general, the protoplasts were stabilized by divalent but not monovalent cations and, unlike whole cells, were sensitive to lysis by Triton X-100. Electron microscopic examination revealed that protoplast formation was preceded by a general breakdown of the cell wall with an apparent squeezing out of the protoplast through the degraded wall. The growth of cells was greatly increased and not accompanied by detectable lysis in a medium modified by elevating the levels of nickel and ammonium.
Acetohydroxy acid synthase is a target for leucine containing peptide toxicity in Escherichia coli
Authors: Gollop N , Tavori H , Barak Z
Published: January 2020

Acetohydroxy acid synthase from a mutant resistant to leucine-containing peptides was insensitive to leucine inhibition. It is concluded that acetohydroxy acid synthase is a target for the toxicity of the high concentrations of leucine brought into Escherichia coli K-12 by leucine-containing peptides.
Polypeptides encoded by the mer operon
Authors: Jackson W J , Summers A O
Published: January 2020

HgCl2-induced polypeptides synthesized by Escherichia coli minicells containing recombinant or natural HgR plasmids were labeled with [35S]methionine and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. All plasmids examined encoded two heavily labeled, HgCl2-inducible polypeptides of 69,000 and 12,000 daltons. Most plasmids also encoded two additional HgCl2-inducible proteins in the 14,000- to 17,000-dalton range. Antiserum prepared against a purified mercuric ion reductase reacts with the 69,000-dalton polypeptide and a minor 66,000-dalton protein seen in several different HgR minicells. Recombinant plasmids constructed from portions of mer DNA from the IncFII plasmid NR1 were also analyzed in the minicell system. Five HgCl2-inducible polypeptides (69,000, 66,000, 15,100, 14,000, and 12,000 daltons) were synthesized in minicells carrying pRR130, a recombinant derivative containing the EcoRI-H and EcoRI-I restriction fragments of NR1. The EcoRI-H fragment of NR1 encodes the three small mer proteins of 15,100, 14,000, and 12,000 daltons and the amino-terminal 40,000 daltons of the mercuric ion reductase monomer.
Alterations in gene expression during heat shock of Achlya ambisexualis
Authors: Gwynne D I , Brandhorst B P
Published: January 2020

When exponentially growing cultures of Achlya ambisexualis strain E87 were raised from their normal growth temperature of 30 degrees C to 35 degrees C, the rates of synthesis of a small number of proteins were dramatically increased. The most predominant proteins synthesized in response to heat shock had molecular weights of 70,000 and 78,000, and their increased synthesis was detected as early as 10 min after the shift to 35 degrees C. Changes in the populations of translatable messenger RNAs during heat shock showed that the levels of the mRNA's for all the major induced proteins correlated very closely with the alterations in the in vivo patterns, suggesting a transcriptional level of control of their synthesis. When after a period of heat shock (60 min) the cultures were shifted back to 30 degrees C, recovery of the preshock patterns of protein synthesis was attained after several hours. Different proteins show temporally distinct patterns of recovery. During recovery the levels of translatable mRNA's for the induced proteins also correlated closely with the patterns of in vivo protein synthesis.
Characterization of factor IIIGLc in catabolite repression-resistant (crr) mutants of Salmonella typhimurium
Authors: Scholte B J , Schuitema A R , Postma P W
Published: January 2020

crr mutants of Salmonella typhimurium are thought to be defective in the regulation of adenylate cyclase and a number of transport systems by the phosphoenolpyruvate-dependent sugar phosphotransferase system, crr mutants are also defective in the enzymatic activity of factor IIIGlc (IIIGlc), a protein component of the phosphotransferase system involved in glucose transport. Therefore, it has been proposed that IIIGlc is the primary effector of phosphotransferase system-mediated regulation of cell metabolism. We characterized crr mutants with respect to the presence and function of IIIGlc by using an immunochemical approach. All of the crr mutants tested had low (0 to 30%) levels of IIIGlc compared with wild-type cells, as determined by rocket immunoelectrophoresis. The IIIGlc isolated from one crr mutant was investigated in more detail and showed abnormal aggregation behavior, which indicated a structural change in the protein. These results supported the hypothesis that a crr mutation directly affects IIIGlc, probably by altering the structural gene of IIIGlc. Several crr strains which appeared to be devoid of IIIGlc in immunoprecipitation assays were still capable of in vitro phosphorylation and transport of methyl alpha-glucoside. This phosphorylation activity was sensitive to specific anti-IIIGlc serum. Moreover, the membranes of crr mutants, as well as those of wild-type cells, contained a protein that reacted strongly with our anti-IIIGlc serum. We propose that S. typhimurium contains a membrane-bound form of IIIGlc which may be involved in phosphotransferase system activity.
Ferrisiderophore reductase activity in Agrobacterium tumefaciens
Authors: Lodge J S , Gaines C G , Arceneaux J E , Byers B R
Published: January 2020

Reduction of the iron in ferriagrobactin by the cytoplasmic fraction of Agrobacterium tumefaciens strictly required NaDH as the reductant. Addition of flavin mononucleotide and anaerobic conditions were necessary for the reaction; when added with flavin mononucleotide, magnesium was stimulatory. This ferrisiderophore reductase activity may be a part of the iron assimilation process in A. tumefaciens.
Carriers in electron transport from molecular hydrogen to oxygen in Rhizobium japonicum bacteroids
Authors: Eisbrenner G , Evans H J
Published: January 2020

An investigation has been conducted to identify electron transport carriers that participate in the oxidation of H2 by H2 uptake-positive strains of Rhizobium japonicum bacteroids. We have observed that the reduced form of dibromothymoquinone at a concentration of 0.2 mM strongly inhibited H2 uptake, endogenous respiration, and C2H2 reduction by bacteroid suspensions. Reduced dibromothymoquinone, however, failed to inhibit the transfer of electrons from H2 to methylene blue under anaerobic conditions, indicating that the hydrogenase per se is insensitive to this inhibitor. Metronidazole, at 1 mM, affected rates of H2 uptake and endogenous respiration only slightly, but strongly inhibited C2H2 reduction. Evidence for H2-dependent cytochrome reduction in an H2 uptake-positive strain of R. japonicum bacteroids is presented. In kinetic studies, the rates of reduction of the type b and c cytochromes in the presence of H2 were shown to be severalfold higher than the rates due to endogenous respiration alone. With hydrogenase-deficient mutants of R. japonicum, no measurable effect of H2 on cytochrome reduction was observed. Our results indicate that ubiquinone and cytochromes of types b and c are involved in the oxyhydrogen reaction in R. japonicum.
MP13, a generalized transducing bacteriophage for Bacillus megaterium
Authors: Vary P S , Garbe J C , Franzen M , Frampton E W
Published: January 2020

The first generalized transducing bacteriophage reported for Bacillus megaterium has been characterized. Optimum conditions for lysate production and transduction procedures were established so that transducing frequencies of 8 x 10(-6) and higher are now possible. The phage, MP13, has a head diameter of 97 nm and a contractile tail (202 by 17 nm) and adsorbs to the periphery of the cell. MP13 was inactivated rapidly at 60 degrees C, but not at 55 degrees C, and was sensitive to toluene, ether, and chloroform. When centrifuged in a neutral CsCl gradient, two bands were observed, a major band of 1.490 g cm-3 and a minor band of 1.482 g cm-3 buoyant density. The major band contained only infective particles, whereas the minor band contained both infective and transducing particles. Phage DNA was resistant to several restriction endonucleases, but yielded 9 fragments with MboI, more than 34 with HindIII, and 7 with BstEII. The molecular weights for the fragments from MboI-BstEII double digests total 97 x 10(9).
Two sites of oxygen control in induced synthesis of respiratory nitrate reductase in Escherichia coli
Authors: Kaprálek F , Jechová E , Otavová M
Published: January 2020

The time course of appearance of respiratory nitrate reductase in Escherichia coli after induction by nitrate was analyzed under different conditions, and the inhibitory effects of oxygen, chloramphenicol, and rifampin were compared. Oxygen appeared to inhibit the synthesis of nitrate reductase at the level of transcription. In addition, the translation or some later steps of enzyme formation were blocked.