bluuebunny/crossref_metadata_2025 huggingface dataset:
Mutation in the structural gene for seryl-transfer ribonucleic acid synthetase of Escherichia coli which affects formation of its gene product at high temperature
Authors: Hill R J , Konigsberg W
Published: January 2020

The rate of formation of seryl-transfer ribonucleic acid (tRNA) synthetase activity was temperature dependent in a temperature-sensitive mutant of Escherichia coli (K28) with an altered seryl-tRNA synthetase structural gene and in a class of spontaneous revertants derived from it. These revertants, which were selected by their ability to grow at 45 degrees C, had high levels of the thermolabile enzyme. The rate of formation of seryl-tRNA synthetase activity in the mutant and in the revertants fell from 100% to near zero with a 4 degrees C temperature range from 40 to 44 degrees C. The temperature-dependent rate of formation of seryl-tRNA synthetase activity was reversible. Dropping the temperature from 44 to 37 degrees C resulted, after a 2- to 3-min delay, in a resumption of the initial rate of formation of enzyme activity. The results could not be accounted for by in vivo or in vitro degradation of the active enzyme. Addition of rifampin just before the temperature shift down from 44 to 37 degrees C inhibited the appearance of seryl-tRNA synthetase activity at the lower temperature. Explanations which might account for these phenomena are proposed.
Shigella toxin inhibition of binding and translation of polyuridylic acid by Escherichia coli ribosomes
Authors: Olenick J G , Wolfe A D
Published: January 2020

Shigella toxin inhibits polyuridylic acid-directed polymerization of phenylalanine in ribosome-enzyme systems obtained from Escherichia coli or from Shigella dysenteriae. The inhibition is the result of toxin acting on ribosomes to prevent polyuridylic acid attachment.
Salmonella typhimurium mutants defective in acetohydroxy acid synthases I and II
Authors: Shaw K J , Berg C M , Sobol T J
Published: January 2020

An analysis of transposon-induced mutants shows that Salmonella typhimurium possesses two major isozymes of acetohydroxy acid synthase, the enzymes which mediate the first common step in isoleucine and valine biosynthesis. A third (minor) acetohydroxy acid synthase is present, but its significance in isoleucine and valine synthesis may be negligible. Mutants defective in acetohydroxy acid synthase II (ilvG::Tn10) require isoleucine, alpha-ketobutyrate, or threonine for growth, a mutant defective in acetohydroxy acid synthase I (ilvB::Tn5) is a prototroph, and a double mutant (ilvG::Tn10 ilvB::Tn5) requires isoleucine plus valine for growth.
Distribution of coenzyme B12-dependent diol dehydratase and glycerol dehydratase in selected genera of Enterobacteriaceae and Propionibacteriaceae
Authors: Toraya T , Kuno S , Fukui S
Published: January 2020

The presence of diol dehydratase and glycerol dehydratase was shown in several bacteria of Enterobacteriaceae grown anaerobically on 1,2-propanediol and on glycerol, respectively. Diol dehydratases of Enterobacteriaceae were immunologically similar, but distinct from that of Propionibacterium freudenreichii.
Rhizobium leguminosarum mutants incapable of normal extracellular polysaccharide production
Authors: Napoli C , Albersheim P
Published: January 2020

Mutants of Rhizobium leguminosarum which are deficient in normal polysaccharide production have been isolated and characterized. A correlation between diminished production of extracellular polysaccharide and reduced infection and nodulation efficiency has been observed.
Beta-2-linked glucans secreted by fast-growing species of Rhizobium
Authors: York W S , McNeil M , Darvill A G , Albersheim P
Published: January 2020

Fast-growing species of Rhizobium were found to secrete low-molecular-weight beta-2-linked glucans when cultured in synthetic liquid medium. These glucans are quite similar to beta-2-linked glucans produced by species of Agrobacterium. No reducing terminus was detected in these glucans.
Identification of a sporulation locus in cloned Bacillus subtilis deoxyribonucleic acid
Authors: Moran C P , Losick R , Sonenshein A L
Published: January 2020

A cloned deoxyribonucleic acid from the purA-cysA region of the Bacillus subtilis chromosome was shown to contain the spoVC locus, a gene whose product is required for sporulation. This is the first demonstration of a spo locus in cloned B. subtilis deoxyribonucleic acid.
uhp-directed, glucose 6-phosphate membrane receptor in Escherichia coli
Authors: Goldenbaum P E , Farmer K S
Published: January 2020

Membrane vesicles were characterized for their ability to specifically bind [14C]glucose 6-phosphate. Membranes prepared from a strain carrying a ColE1 uhp hybrid plasmid showed significantly enhanced glucose 6-phosphate binding. It is hypothesized that glucose 6-phosphate binding to these membranes is due to a uhpR-directed, membrane-bound receptor which functions in regulation of the inducible uhpT gene product: the hexose phosphate permease.
On the question of chromosomal gene transfer via conjugation in Neisseria gonorrhoeae
Authors: Steinberg V I , Goldberg I D
Published: January 2020

Transfer of chromosomal markers between cells of Neisseria gonorrhoeae does not require the presence of a 24.5-megadalton conjgal plasmid in the donor. Apparent conjugal transfer of chromosomal markers may be the result of leakage of deoxyribonucleic acid by some cells in the mating mixture and subsequent uptake of this deoxyribonucleic acid by others.
Methylation of ribosomal proteins in Bacillus subtilis
Authors: Mardones E , Amaro A M , Jerez C A
Published: January 2020

We measured the methylation of ribosomal proteins from the 30S and 50S subunits of Bacillus subtilis after growing the cells in the presence of [1-14C]methionine and [methyl-3H]methionine. Two-dimensional polyacrylamide gel electrophoretic analysis revealed a preferential methylation of the 50S ribosomal proteins. Proteins L11 and L16, and possibly L9, L10, L18, and L20, were methylated. On the other hand, only two possibly methylated proteins were found on the 30S subunit. A comparison of these results with those for Escherichia coli suggests a common methylation pattern for the bacterial ribosomal proteins.
Regulation of nitrogen metabolism in glutamine auxotrophs of Klebsiella pneumoniae
Authors: Leonardo J M , Goldberg R B
Published: January 2020

We examined the regulation of nitrogen metabolism in four classes (glnA, glnB, glnF, and glnG) of Gln- auxotrophs of Klebsiella pneumoniae. These studies indicate that glutamine synthetase does not directly mediate the physiological response to NH4+ in this organism. We present evidence suggesting that the effect of NH4+ on the expression of genes involved in nitrogen metabolism involves the products of the glnF and glnG genes.
Regulation of lysine- and lysine-plus-threonine-inhibitable aspartokinases in Bacillus brevis
Authors: Hitchcock M J , Hodgson B , Linforth J L
Published: January 2020

Further studies on the expression of the two aspartokinase activities in Bacillus bovis are presented. Aspartokinase I (previously shown to be inhibited and repressed by lysine) was found to be repressed by diaminopimelate in the wild-type strain. However, in a mutant unable to convert diaminopimelate to lysine, starvation for lysine resulted in an increase in aspartokinase I activity. Thus, lysine itself or an immediate metabolite was the true effector of repression. Aspartokinase II (previously shown to be inhibited by lysine plus threonine) was repressed by threonine. Studies with the parent strain and auxotrophs inidicated that only threonine or an immediate metabolite of threonine was involved in this repression. Methionine and isoleucine were not effectors of any of the detected aspartokinase activities. Apart from inhibition and repression controls, a third as yet undefined regulatory mechanism operated to decrease the levels of both aspartokinases as growth declined, even in mutants in which repression control was absent. In thiosine-resistant, lysine-excreting mutants with elevated levels of aspartokinase, the increase in activity could always be attributed to one enzyme or the other, never both. The existence of separate structural genes for each aspartokinase is therefore suggested.
Physiological basis for meso-tartrate sensitivity in some strains of Salmonella typhimurium
Authors: Old D C , Alfredsson G A , Brown C M
Published: January 2020

meso-Tartrate inhibited the growth of non-meso-tartrate-utilizing strains of Salmonella typhimurium in peptone water media and mineral salts media with some, but not all, carbon sources. C-R intermediates of the tricarboxylic acid cycle or compounds readily converted to them and substrates metabolized independently of the C-6 part of the cycle spared bacteria from the inhibitory effects of meso-tartrate when added to cultures along with meso-tartrate. Experiments with cell-free extracts of non-meso-tartrate-utilizing strains from batch and continuous cultures showed that meso-tartrate was a competitive inhibitor of isocitrate dehydrogenase and isocitrate lyase activities and also inhibited citrate synthase and malate synthase activities. The synthesis of these enzymes was not inhibited by meso-tartrate. The isocitrate enzymes of meso-tartrate-utilizing strains of S. typhimurium were similarly inhibited by meso-tartrate, but inhibition of the growth of meso-tartrate-utilizing strains was demonstrable only in uninduced cultures in which the intracellular concentrations of meso-tartrate were high.
Lambda Receptor in the Outer Membrane of
Escherichia coli
as a Binding Protein for Maltodextrins and Starch Polysaccharides
Authors: Ferenci Thomas , Schwentorat Marina , Ullrich Susanne , Vilmart Jeannine
Published: January 2020

The starch polysaccharides amylose and amylopectin are not utilized by
Escherichia coli
, but are bound by the bacteria. The following evidence supports the view that the outer membrane λ receptor protein, a component of the maltose/ maltodextrin transport system is responsible for the binding. (i) Amylose and amylopectin both inhibit the transport of maltose into
E. coli
. (ii) Both polysaccharides prevent binding of non-utilizable maltodextrins by the intact bacterium, a process previously shown to be dependent on components of the maltose transport system (T. Ferenci, Eur. J. Biochem., in press). (iii) A fluorescent amylopectin derivative,
O
-(fluoresceinyl thiocarbamoyl)-amylopectin, has been synthesized and shown to bind to
E. coli
in a reversible, saturable manner. Binding of
O
-(fluoresceinyl thiocarbamoyl)-amylopectin is absent in mutants lacking the λ receptor, but mutations in any of the other components of the maltose transport system do not affect binding as long as λ receptor is present. (iv) Using the inhibition of λ receptor-dependent
O
-(fluoresceinyl thiocarbamoyl)-amylopectin binding as an assay, the affinities of the λ receptor for maltodextrins and other sugars have been estimated. The affinity for dextrins increases with increasing degree of polymerization (

K
d

for maltose, 14 mM; for maltotetraose, 0.3 mM; for maltodecaose, 0.075 mM). Maltose and some other di- and trisaccharides are inhibitory to amylopectin binding, but only at concentrations above 1 mM.
Interactions of competent Streptococcus sanguis (Wicky) cells with native or denatured, homologous or heterologous deoxyribonucleic acids
Authors: Ranhand J M
Published: January 2020

Competent cell-deoxyribonucleic acid (DNA) interactions were examined using tritium-labeled homologous or heterologous native or denatured DNAs and competent Streptococcus sanguis Wicky cells (strain WE4). The DNAs used were extracted from WE4 cells, Escherichia coli B cells, and E. coli bacteriophages T2, T4, T6, and T7. The reactions examined were: (i) total DNA binding, (ii) deoxyribonuclease-resistant DNA binding, and (iii) the production of acid-soluble products from the DNA. Optimal temperatures for the reactions were as follows: reaction (i), between 30 and 40 degrees C; reaction (ii), 30 degrees C; and reaction (iii), greater than 40 degrees C. The rates for the reactions (expressed as molecules of DNA that reacted per minute per colony-forming unit) did not vary greatly from one DNA source to another. With a constant competent cell concentration and differing DNA concentrations below a saturation level (from a given source), a different but constant fraction of the added DNA was cell bound, deoxyribonuclease resistant, and degraded to acid-soluble products. In experiments where the number of competent cells was varied and the DNA concentration was held constant, again essentially the same result was obtained. The extent of reactions (i), (ii), and (iii) depended upon the numbers as well as the source of DNA molecules applied to competent cells. Calcium ion essential for native DNA-cell reactions was also found essential for denatured DNA-cell reactions. Data obtained from competition experiments lead to the conclusion that competent WE4 cells contain specific sites for native as well as denatured DNAs.
Properties of Haemophilus influenzae mutants that are slightly recombination deficient and carry a mutation in the rec-1 gene region
Authors: Kooistra J , Venema G
Published: January 2020

The highly recombination-deficient rec-1 mutants of Haemophilus influenzae are, as far as tested, equivalent to recA mutants of Escherichia coli. By selection for mutations in the rec-1 gene of H. influenzae, mutants designated ird (intermediary recombination-deficient) mutants were isolated; these mutants were much less recombination deficient (degree of transformability, 0.2 to 30% of wild-type value) than previously isolated rec-1 mutants (degree of transformability, 0.0001% of wild-type value). The ird mutants were more sensitive to ultraviolet irradiation and mytomycin C treatment than the wild type, but less sensitive than rec-1 mutants. Spontaneous production of phage HP1c1 by lysogenic MC11 cells and prophage induction by mitomycin C or ultraviolet irradiation were the same as in the wild type. In the ird mutants endogenous deoxyribonucleic acid was degraded both spontaneously and after ultraviolet irradiation to the same extent as in the wild type. Examination of one of the ird mutants revealed that recombination could be enhanced by ultraviolet irradiation, possibly because of an increased synthesis of the rec-1 gene product induced by ultraviolet irradiation.
Production of methanol from aromatic acids by Pseudomonas putida
Authors: Donnelly M I , Dagley S
Published: January 2020

When grown at the expense of 3,4,5-trimethoxybenzoic acid, a strain of Pseudomonas putida oxidized this compound and also 3,5-dimethoxy-4-hydroxybenzoic (syringic) and 3,4-dihydroxy-5-methoxybenzoic (3-O-methylgallic) acids; but other hydroxy- or methoxy-benzoic acids were oxidized slowly or not at all. Radioactivity appeared exclusively in carbon dioxide when cells were incubated with [4-methoxyl-14C]trimethoxybenzoic acid, but was found mainly in methanol when[methoxyl-14C]3-O-methylgallic acid was metabolized. The identity of methanol was proved by analyzing the product from [methoxyl-13C]3-O-methylgallic acid by nuclear magnetic resonance spectroscopy and by isolating the labeled 3,5-dinitrobenzoic acid methyl ester, which was examined by mass spectrometry. These results, together with measurements of oxygen consumed in demethylations catalyzed by cell extracts, showed that two methoxyl groups of 3,4,5-trimethoxybenzoate and one of syringate were oxidized to give carbon dioxide and 3-O-methylgallate. This was then metabolized to pyruvate; the other product was presumed to be the 4-methyl ester of oxalacetic acid, for which cell extracts contained an inducible, specific esterase. P. putida did not metabolize the methanol released from this compound by hydrolysis. Support for the proposed reaction sequence was obtained by isolating mutants which, although able to convert 3,4,5-trimethoxybenzoic acid into 3-O-methylgallic acid, were unable to use either compound for growth.
Outer Membrane-Dependent Transport Systems in
Escherichia coli
: Effect of Repression or Cessation of Colicin Receptor Synthesis on Colicin Receptor Activities
Authors: Kadner Robert J. , McElhaney Gail
Published: January 2020

Proteins in the outer membrane of gram-negative bacteria serve as general porins or as receptors for specific nutrient transport systems. Many of these proteins are also used as receptors initiating the processes of colicin or phage binding and uptake. The functional activities of several outer membrane proteins in
Escherichia coli
K-12 were followed after cessation or repression of their synthesis. Cessation of receptor synthesis was accomplished with a thermolabile suppressor activity acting on amber mutations in
btuB
(encoding the receptor for vitamin B
12
, the E colicins, and phage BF23) and in
fepA
(encoding the receptor for ferric enterochelin and colicins B and D). After cessation of receptor synthesis, cells rapidly became insensitive to the colicins using that receptor. Treatment with spectinomycin or rifampin blocked appearance of insensitive cells and even increased susceptibility to colicin E1. Insensitivity to phage BF23 appeared only after a lag of about one division time, and the receptors remained functional for B
12
uptake throughout. Therefore, possession of receptor is insufficient for colicin sensitivity, and some interaction of receptor with subsequent uptake components is indicated. Another example of physiological alteration of colicin sensitivity is the protection against many of the
tonB
-dependent colicins afforded by provision of iron-supplying siderophores. The rate of acquisition of this nonspecific protection was found to be consistent with the repression of receptor synthesis, rather than through direct and immediate effects on the
tonB
product or other components of colicin uptake or action.
Inositol-Limited Growth, Repair, and Translocation in an Inositol-Requiring Mutant of
Neurospora crassa
Authors: Hanson Barbara A.
Published: January 2020

The biochemical consequences of inositol limitation in an inositol auxotroph of
Neurospora crassa
have been examined as a means of disclosing the cellular role of inositol. The cellular levels of inositol in the
inl
mutant were proportional to the concentration of inositol in the growth medium whereas inositol phosphate levels remained relatively constant at about 0.1 μmol/g (dry weight). After 72 h of growth, about 57-fold more protein per milligram (dry weight) was released by the mutant grown on limiting inositol than by the inositol-supplemented control. When the inositol-limited growth medium was osmotically buffered with 1% NaCl, 3% NaCl, or 6% sorbitol, there was about 33, 74, or 54%, respectively, less protein released by the mutant. These results are consistent with cell lysis occurring in the mutant grown on limiting inositol because of a structurally weakened cell wall and membrane deterioration. When sufficient inositol for normal mycelial growth was supplied to an inositol-deficient mycelium, there was within 2 h a rapid incorporation of inositol to 85% of control levels. This incorporation occurred without significant growth by any area of the mycelium. About 10 to 15% of the total cell inositol was translocated forward from the older mycelial areas to the growing tips; only 2 to 5% of the total cell inositol was translocated backward toward the older mycelial areas. Possible mechanisms of translocation are discussed.
Suppressors of a UGG missense mutation in Escherichia coli
Authors: Murgola E J , Childress J R
Published: January 2020

As part of our investigation of tRNA structure-function relationships, we isolated and preliminarily characterized translational suppressors of the tryptophan codon UGG in a trpA missense mutant of Escherichia coli. the parent strain also contained two other mutant alleles relevant to the suppressor search; these were supD, which codes for a serine-inserting amber suppressor tRNA, and gly V55, the gene for a GGA/G-reading mutationally altered glycine tRNA. On the basis of map location, reversed-phase (RPC-5) column chromatography of glycyl-tRNA, and codon response, several classes have been distinguished so far. The number of suppressors in each class, their codon responses, and their apparent genic identities, respectively, are as follows: class 1--4 suppressors, UGG, supD; class 2--12 suppressors, UGG, glyU; class 3--9 suppressors, UGA and UGG, glyT; class 4--2 suppressors, UGG, glyT; class 5--7 suppressors, UGG, gly V55. Besides these, one suppressor retains supD activity, but so far its map location has not been distinguished from that of supD. Another suppressor clearly does not map near supD or any of the glycine tRNA genes mentioned. These last two suppressors may represent novel missense suppressors such as misacylated tRNA's or mutationally altered aminoacyl-tRNA synthetases, tRNA modification enzymes, or ribosomes. Finally, three other suppressors were obtained from a strain containing glyT56, the gene for an AGA/G-reading form of glyT tRNA. All three occurred at the expense of glyT56 activity and exhibited the the transductional linkage to argH that is characteristic of glyT.
Biosynthesis of
d
-Alanyl-Lipoteichoic Acid: Characterization of Ester-Linked
d
-Alanine in the In Vitro-Synthesized Product
Authors: Childs Warren C. , Neuhaus Francis C.
Published: January 2020

d
-Alanyl-lipoteichoic acid (
d
-alanyl-LTA) contains
d
-alanine ester residues which control the ability of this polyer to chelate Mg
2+
. In
Lactobacillus casei
a two-step in vitro reaction sequence catalyzed by the
d
-alanine-activating enzyme and
d
-alanine:membrane acceptor ligase incorporates
d
-alanine into membrane acceptor. In this paper we provide additional evidence that the in vitro system catalyzes the covalent incorporation of
d
-[
14
C]alanine into membrane acceptor which is the poly([
3
H]glycerol phosphate) moiety of
d
-alanyl-LTA. This conclusion was supported by the observation that the
d
-[
14
C]alanine and [
3
H]glycerol labels of the partially purified product were co-precipitated by antiserum containing globulins specific for poly(glycerol phosphate). The isolation of
d
-[
14
C]alanyl-[
3
H]glycerol from
d
-[
14
C]alanine·[
3
H]glycerol-labeled
d
-alanyl-LTA synthesized in the in vitro system indicated that the
d
-alanine was linked to the poly(glycerol phosphate) chain of the LTA. A comparison of the reactivities of the
d
-alanine residues of
d
-alanyl-glycerol and
d
-alanyl-LTA supported the conclusion that the incorporated residue of
d
-alanine was attached by an ester linkage. Thus, the data indicated that the in vitro system catalyzes the incorporation of
d
-alanine covalently linked by ester linkages to the glycerol moieties of the poly(glycerol phosphate) chains of
d
-alanyl-LTA. New procedures are presented for the partial purification of
d
-alanyl-LTA with a high yield of ester-linked
d
-alanine and for the sequential degradation of the poly(glycerol phosphate) moiety substituted with
d
-alanine of
d
-alanyl-LTA with phosphodiesterase II/phosphatase from
Aspergillus niger
.
Influence of chromosome integrity on Escherichia coli cell division
Authors: Markham B E , Brubaker R R
Published: January 2020

The antitumor agent cis-platinum(II)diamminodichloride (PDD) caused wild-type and recA+ deoxyribonucleic acid (DNA) repair-deficient mutant cells of Escherichia coli K-12 to grow as long, multinucleated filaments. At 5 micrograms/ml, the times required for reduction of viability to 37% for wild-type, polA, recB,C, uvrA, and recA organisms were > 200, 200, 120, 25, and 5 min, respectively. Only recA cells exhibited @reckless" degradation of DNA at this concentration of PDD. As shown by sedimentation in alkaline sucrose gradients, generation of single-strand breaks in DNA of the remaining organisms was a major consequence of growth in PDD. Upon incubation in fresh medium after removal of the compound and storage for 4 h at 4 degrees C, a respective lag of 3, 4, 6, and 9 h occurred before filaments of wild-type, polA, recB,C, and uvrA cells commenced cell division. Maintenance at 4 degrees C, which evidently delayed postshift initiation of chromosome replication, was only essential for fragmentation of uvrA filaments. In all cases, these periods of division delay corresponded to those required for restoration of normal chromosomal molecular weight as determined in alkaline sucrose gradients.
Distribution of calcium and other elements in cryosectioned Bacillus cereus T spores, determined by high-resolution scanning electron probe x-ray microanalysis
Authors: Stewart M , Somlyo A P , Somlyo A V , Shuman H , Lindsay J A , Murrell W G
Published: January 2020

The distribution of a number of key elements in Bacillus cereus T spores was determined by high-resolution scanning electron probe X-ray microanalysis. To circumvent the redistribution of soluble or weakly bound elements, freeze-dried cryosections of spores, which had been rapidly frozen in 50% aqueous polyvinyl pyrrolidone, were employed. The sections were examined by using a modified Philips EM400 electron microscope fitted with a field emission gun, scanning transmission electron microscopy attachment, and a computer-linked energy-dispersive X-ray microanalysis system. X-ray maps for selected elements and the corresponding electron image were produced simultaneously by scanning the cryosections with a fine electron beam in a raster pattern, using the scanning transmission electron microscopy attachment. The results indicated that almost all of the calcium, magnesium, and manganese, together with most of the phosphorus, was located in the core region. An unexpectedly high concentration of silicon was found in the cortex/coat layer. Granules containing high concentrations of calcium, manganese, and phosphorus were demonstrated in spores containing reduced levels of dipicolinic acid. Spot mode analyses, in which a stationary beam was located over the region of interest in the spore cryosection, confirmed the results obtained with the scanning mode and also provided a more accurate quantitation of the elemental concentrations on a dry weight bases.
Dimer excision in Escherichia coli in the presence of caffeine
Authors: Rothman R H
Published: January 2020

The observation that polA1 and recL152 mutations result in both slow pyrimidine dimer excision and large repair patch size leads to the hypothesis that patch size is directly related to the rate of excision. In this study caffeine, a known inhibitor of excision repair, was used to examine the extent of correlation between excision rate and patch size by measuring patch size in the presence of several concentrations of caffeine. Both the rate of excision and the resistance to ultraviolet radiation were reduced with increasing concentrations of caffeine after irradiation. Caffeine also inhibited the rate at which incisions were made and prolonged the time required to rejoin the discontinuities. Patch size, however, was unaffected by caffeine treatment.
Functional analysis of minichromosome replication: bidirectional and unidirectional replication from the Escherichia coli replication origin, oriC
Authors: Meijer M , Messer W
Published: January 2020

Replicating molecules of minichromosomes pCM959 and pOC24 were analyzed by electron microscopy. Replication of pCM959 proceeded bidirectionally from the replication origin, oriC, in about 60% of the molecules; the rest of the molecules replicated unidirectionally in either direction. pOC24, in which deoxyribonucleic acid to the right (clockwise) of the oriC segment is deleted, seemed to replicate predominantly unidirectionally counterclockwise from oriC.
pKJ1, a naturally occurring conjugative plasmid coding for toluene degradation and resistance to streptomycin and sulfonamides
Authors: Yano K , Nishi T
Published: January 2020

Pseudomonas sp. TA8 isolated by m-toluate enrichment from an aqueous sample metabolized toluene and m- and p-xylenes via the meta cleavage pathway, and manifested specific resistance to streptomycin and sulfonamides. A variety of experiments revealed that the pKJ1 plasmid of about 150 megadaltons carried by TA8 specified both the toluene and xylene degradative function (the Tol function) and streptomycin/sulfonamide resistance. The deletion of a segment of pKJ1 (about 22 megadaltons) resulted in the loss of the Tol function. pKJ1 was not assigned to Pseudomonas incompatibility group P-1, P-2, P-3, or P-9.
Patterns of polarity in the Escherichia coli car AB gene cluster
Authors: Gigot D , Crabeel M , Feller A , Charlier D , Lissens W , Glansdorff N , Piérard A
Published: January 2020

The direction of transcription of the carAB gene cluster, which codes for Escherichia coli carbamoylphosphate synthase, was deduced from the effects of phage Mu-1 insertions in each of the two genes and from the results of ribonucleic acid-deoxyribonucleic acid hybridization experiments relating the quantity of car messenger ribonucleic acid to the location of various car mutations. The car locus appears to constitute an operon polarized from carA to carB. The levels of carA and carB products were determined in a large number of car mutants by using in vitro and in vivo complementation assays. The results obtained display strong anomalies, which are discussed in light of the conclusions described above.
Regulation of Escherichia coli K-12 hexuronate system genes: exu regulon
Authors: Portalier R , Robert-Baudouy J , Stoeber F
Published: January 2020

Two types of Escherichia coli K-12 regulatory mutants, partially or totally negative for the induction of the five catabolic enzymes (uronic isomerase, uxaC; altronate oxidized nicotinamide adenine dinucleotide: uxaB; mannonate hydrolyase, uxuA) and the transport system (exuT) of the hexuronate-inducible pathway, were isolated and analyzed enzymatically. Hexuronate-catabolizing revertants of the negative mutants showed a constitutive synthesis for some or all of these enzymes. Negative and constitutive mutations were localized in the same genetic locus, called exuR, and the following order for the markers situated between the min 65 and 68 was determined: argG--exuR--exuT--uxaC--uxaA--tolC. The enzymatic characterization of the pleiotropic negative and constitutive mutants of the exuR gene suggests that the exuR regulatory gene product exerts a specific and total control on the three exuT, uszB, and uxaC-uxaA operons of the galacturonate pathway and a partial control on the uxuA-uxuB operon of the glucuronate pathway. The analysis of diploid strains conatining both the wild type and a negative or constitutive allele of the exuR gene, as well as the analysis of thermosensitive mutants of the exuR gene, was in agreement with a negative regulatory mechanism for the control of the hexuronate system.
Covalently closed circular deoxyribonucleic acids in spiroplasmas
Authors: Ranhand J M , Mitchell W O , Popkin T J , Cole R M
Published: January 2020

Ten of twelve spiroplasma strains from different sources carried multiple covalently closed circular duplex deoxyribonucleic acid molecules, as shown by ethidium bromide-cesium chloride gradient centrifugation of cell lysates and examination of resulting bands by electron microscopy and agarose gel electrophoresis. Two to eight size classes per strain, comprising molecules of masses from 1 X 10(6) to 26 X 10(6), were detected. Several size classes of molecules were found in common in different spiroplasma strains. The amount of covalently closed circular deoxyribonucleic acid per strain was as much as 12% of total cellular deoxyribonucleic acid. The presence of sizes of the circular molecules appeared unrelated to either carriage or active production of known spiroplasma viruses, and it is tentatively concluded that they are plasmids rather than genomes or replicative forms of viruses.
tif-dependent induction of colicin E1, prophage lambda, and filamentation in Escherichia coli K-12
Authors: Tessman E S , Peterson P K
Published: January 2020

To help understand how the tif-1 mutation of the recA gene of Escherichia coli confers adenine activability on the recA protein, we used the fact that cytidine plus guanosine inhibits induction of prophage lambda and cell filamentation in a tif-1 mutant, and that adenine reverses this inhibition. We varied the amount of adenine in agar plates containing a fixed amount of cytidine and scored for survivors of three different tif-dependent lethal induction processes. Much more adenine was required for cell killing when cytidine was present than when it was absent. Therefore adenine does not override cytidine inhibition, but instead appears to compete with it for a site of action which may be on the recA protein. The competition is not at the cell transport level. Our results lead to a model in which the tif form of the recA protein is an allosteric enzyme that binds both negative and positive modulators. By varying the adenine-cytidine ratio of the medium it is possible to control the degree of induction in a tif-1 cell. For the three different tif-dependent inductions studied here, least adenine was required for lambda induction and most for lethal filamentation, presumably reflecting requirements for different amounts of activated recA protein in each process. Varying the adenine-cytidine ratio revealed two stable intermediate stages in lambda induction, as well as a stage of colicin E1 induction in which the cells produced colicin without cell death. The rate of filament formation could be similarly controlled. Experiments with tif (ColE1, lambda) gave evidence of a competition between colicin repressor and lambda repressor for activated recA protein.
Isolation of ara-lac gene fusions in Salmonella typhimurium LT2 by using transducing bacteriophage Mu d (Apr lac)
Authors: Lee J H , Heffernan L , Wilcox G
Published: January 2020

A specialized Mu transducing phage containing a gene encoding ampicillin resistance and the lac structural genes without the lac promotor [Mu d(apr lac)] has been constructed and used to create gene fusions in Escherichia coli (M. J. Cadadaban and S. N. Cohen, Proc. Natl. Acad. Sci. U.S.A. 76:4530--4533, 1979). Transposition of the Mu d(Apr lac) phage to chromosomal sites can result in lac expression being controlled by a chromosomal promoter. We have constructed an Escherichia coli K-12 strain in which the Mu d(Apr lac) phage is integrated into an F factor. The F+::Mu d(Apr lac) was then transferred by conjugation into a Salmonella typhimurium strain that was sensitive to L-arabinose. Strains containing gene fusions were selected as L-arabinose-resistant colonies after partial induction of the phage. Two classes of ara-lac fusion strains were isolated: (i) araC-lac fusions in which the expression of beta-galactosidase synthesis was constitutuve and not inducible by L-arabinose; and ((ii) fusion of the lac genes to the ara structural genes in which the expression of beta-galatosidase synthesis was induced 263-fold by L-arabinose.
Substrate channeling: alpha-ketobutyrate inhibition of acetohydroxy acid synthase in Salmonella typhimurium
Authors: Shaw K J , Berg C M
Published: January 2020

Excess alpha-ketobutyrate inhibited the growth of Salmonella typhimurium LT2 by inhibiting the acetohydroxy acid synthase-catalyzed synthesis of alpha-acetolactate (a valine precursor). As a result, cells were starved for valine, and both ilvB (encoding acetohydroxy acid synthase I) and ilvGEDA (ilvG encodes acetohydroxy acid synthase II) were derepressed. The addition of valine reversed the effects of alpha-ketobutyrate.
Acetaldehyde coenzyme A dehydrogenase of Escherichia coli
Authors: Clark D P , Cronan J E
Published: January 2020

Mutants of Escherichia coli (adh) in which alcohol dehydrogenase is derepressed under aerobic conditions were also found to overproduce acetaldehyde coenzyme a dehydrogenase. However, acetaldehyde coenzyme A dehydrogenase was induced by ethanol or acetaldehyde and subject to strong catabolite repression, whereas alcohol dehydrogenase was little affected by these conditions. Mutants no longer able to use ethanol as carbon source were isolated from an adh strain. Some of these mutants were revertants at the adh locus and no longer produced either alcohol dehydrogenase or acetaldehyde coenzyme A dehydrogenase. Others, designated acd, were found to lack only acetaldehyde coenzyme A dehydrogenase. The acd mutation was located at min 62 of the E. coli genetic map, the gene order being thyA-lysA-acd-serA-fda. Isolation of Tn10 insertions cotransducible with acd greatly simplified the mapping procedure.
Morphogenesis of Candida albicans and cytoplasmic proteins associated with differences in morphology, strain, or temperature
Authors: Manning M , Mitchell T G
Published: January 2020

The extent of change in cytoplasmic proteins which accompanies yeast-to-mycelium morphogenesis of Candida albicans was analyzed by two-dimensional gel electrophoresis. Pure cultures of yeasts and true hyphae (i.e., without concomitant production of pseudohyphae) were grown in a synthetic low-sulfate medium. The two strains selected for this study were strain 4918, which produces pure mycelial cultures in low-sulfate medium at 37 degrees C and yeast cells at 24 degrees C, and strain 2252, which produces yeast cells exclusively at both 24 and 37 degrees C in low-sulfate medium. The proteins of both strains were labeled at both temperatures with [35S]sulfate, cytoplasmic fractions were prepared by mechanical disruption and ultracentrifugation, and the labeled proteins were analyzed by two-dimensional electrophoresis. Highly reproducible protein spot patterns were obtained which defined hundreds of proteins in each extract. Ten protein spots were identified on the two-dimensional gels of the 4918 mycelial-phase extract which were not present in the 4918 yeast-phase extract. These proteins appeared to be modifications of preexisting yeast-phase proteins rather than proteins synthesized de novo in the mycelial cells because 5 were absorbed by rabbit anti-yeast-phase immunoglobulin and each of the 10 was also present in extracts of strain 2252 grown at 24 and 37 degrees C, indicating that they were neither unique to filamentous cells nor sufficient for induction or maintenance of the mycelial morphology. Thirty-three proteins were identified in the 4918 yeast-phase extract which were not present in the 4918 mycelial-phase extract. Pulse-chase experiments revealed the synthesis of new proteins during yeast-to-mycelial conversion, but none of these was unique to mycelial cells. No differences in the major cytoplasmic proteins of any of the yeast- or mycelial-phase extracts were identified. This finding suggests that the major structural proteins of the cytoplasm are not extensively modified and argues instead that proteins unique to either phase may serve a regulatory function.
Intracellular structures of Mycoplasma pneumoniae revealed after membrane removal
Authors: Meng K E , Pfister R M
Published: January 2020

Mycoplasma pneumoniae was grown on Formvar- and carbon-coated electron microscope grids and treated with the nonionic detergent Triton X-100 to gently remove the membrane and cytoplasm. The detergent mixture was composed of 0.5% Triton X-100 in SSR-2 broth base. After this treatment, the grids were rinsed in a mixture of 0.1 M KCl, 5 mM MgCl2, and 6 mM potassium phosphate buffer (pH 7.05) and negatively stained with uranyl acetate. The Triton X-100-resistant remains of M. pneumoniae after gentle removal of the membrane and cytoplasm consisted of fibrous structures oriented similarly to the undisrupted cells. The thin fibers displayed a negative staining quality and diameter analogous to that of rabbit muscle F-actin. The fibrous moieties ended in rodlike condensations which appeared striated in negatively stained and shadowed preparations. These striations were regular, and the majority of rod structures had lengths of 220 to 300 nm and widths of 50 to 80 nm. Specific antibody to rabbit muscle actin, produced in guinea pigs, was used in indirect immunofluorescence of the M. pneumoniae colonies. Fluorescence was detected, with concentrations at the colony center and at the tips of filamentous cells.
Identification of new genes in a cell envelope-cell division gene cluster of Escherichia coli: cell division gene ftsQ
Authors: Begg K J , Hatfull G F , Donachie W D
Published: January 2020

We report the identification, cloning, and mapping of a new cell division gene, ftsQ. This gene formed part of a cluster of three division genes (in the order ftsQ ftsA ftsZ) which itself formed part of a larger cluster of at least 10 genes, all of which were involved in some step in cell division, cell envelope synthesis, or both. The ftsQAZ group was transcribed from at least two independent promoters.
Characterization of type 1 pili of Salmonella typhimurium LT2
Authors: Korhonen T K , Lounatmaa K , Ranta H , Kuusi N
Published: January 2020

Type 1 pili from Salmonella typhimurium LT2 were purified and characterized. The pilus filaments were 6 nm in diameter and over 1 microns long. Estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight of the pilin was 21,000. The isoelectric point of the filament was 4.1. Hydrophobic amino acids comprised 40.3% of the total amino acids of the pilin, which contained more proline, serine, and lysine than reported for the type 1 pilin of Escherichia coli. Purified pili agglutinated both horse and chicken erythrocytes and yeast cells but not bovine, sheep, or human erythrocytes. Horse erythrocyte agglutination was inhibited at lower concentrations by alpha-methyl-D-mannoside than by yeast mannane and D-fructose. Agglutination was not affected by D-galactose or sucrose. Results of the present study confirm the role of type 1 pili as Salmonella hemagglutinins and show chemical differences between the type 1 pili of S. typhimurium and E. coli.
Peptidoglycolipid nature of the superficial cell wall sheath of smooth-colony-forming mycobacteria
Authors: Barrow W W , Ullom B P , Brennan P J
Published: January 2020

The most superficial cell wall layer present in smooth-colony-forming mycobacteria was isolated from serovar 20 of the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum (MAIS) serocomplex and examined chemically and by electron microscopy. Most (70 to 80%) of the fibrillar material consisted of an array of serologically active, acetylated C-myosidic peptidoglycoplipids with the basic structure (formula, see text) but in which the location of acetyl groups and the arrangement of monosaccharides have not been defined. Apparently, all serovars within the MAIS complex are characterized by structurally related superficies in which the monoglycosyl-lipopeptide portion is invariable but the oligosaccharide attachment is peculiar to each serovar. These unique inert structures may be an important factor in shielding the pathogen within phagolysosomes from lysosomal enzymes.
Photorepair of ultraviolet-induced petite mutational damage in Saccharomyces cerevisiae requires the product of the PHR1 gene
Authors: Green G , MacQuillan A M
Published: January 2020

A wild-type (phr+) diploid yeast strain showed photorepair of petite mutational damage, whereas a photoreactivation-deficient (phr1/phr1) diploid strain did not, indicating that the PHR1 gene product was required for mitochondrial photorepair.
Increase of Nitrogenase Activity in the Blue-Green Alga
Nostoc muscorum
(Cyanobacterium)
Authors: Scherer Siegfried , Kerfin Wolfgang , Böger Peter
Published: January 2020

Preincubation of the blue-green alga (cyanobacterium)
Nostoc muscorum
under hydrogen or argon (nongrowing conditions, neither CO
2
nor N
2
or bound nitrogen present) in the light resulted in a two- to fourfold increase of light-induced hydrogen evolution and a 30% increase of acetylene reduction. Preincubation under the same gases in the dark led to a decrease of both activities. Cultivation of algae under a hydrogen-containing atmosphere (N
2
, H
2
, CO
2
) increased neither hydrogen nor ethylene evolution by the cells. Formation of both ethylene and hydrogen is due to nitrogenase activity, which apparently was induced by the absence of N
2
or bound nitrogen and not by the presence of hydrogen. Inhibitors of protein biosynthesis prevented the increase of nitrogenase activity. Hydrogen uptake by the cells was almost unaffected under all of these conditions. With either ammonia or chloramphenicol present, nitrogenase activity decreased under growing conditions (i.e., an atmosphere of N
2
and CO
2
). The kinetics of decrease were the same with ammonia or chloramphenicol, which was interpreted as being due to rapid protein breakdown with a half-life of approximately 4 h. The decay of nitrogenase activity caused by chloramphenicol could be counteracted by nitrogenase-inducing conditions, i.e., by the absence of N
2
or bound nitrogen. A cell-free system from preconditioned algae with an adenosine 5′-triphosphate-generating system exhibited the same increase or decrease of nitrogenase activity as the intact cell filaments, indicating that this effect resided in the nitrogenase complex only. We tentatively assume that not the whole nitrogenase complex, but merely a subunit or a special protein with regulatory function, is susceptible to fast turnover.
Bacteriophage infection interferes with guanosine 3'-diphosphate-5'-diphosphate accumulation induced by energy and nitrogen starvation in the cyanobacterium Anacystis nidulans
Authors: Borbély G , Kaki C , Gulyás A , Farkas G L
Published: January 2020

Anacystis nidulans accumulates large amounts of guanosine 3'-diphosphate-5'-diphosphate (ppGpp) upon nutritional or energy starvation induced by light-to-dark shift, treatment with carbonylcyanide-m-chlorophenylhydrazone (an uncoupler), or treatment with L-methionine-DL-sulfoximine (an inducer of nitrogen starvation). In contrast to healthy A. nidulans cells, those infected by AS-1 cyanophage do not respond with ppGpp accumulation when starved after about one-third of the complete infection cycle, except, to some extent, under extreme conditions when both nitrogen deprivation and energy deprivation are induced simultaneously (darkening plus L-methionine-DL-sulfoximine treatment). In contrast to cyanophage infection in Anacystis, infection with T4 phage of Escherichia coli CP 78 cells does not affect their accumulation of ppGpp under treatments identical with or similar to those applied in the experiments with Anacystis. This difference in response of phage-infected heterotrophic and photoautotrophic cells to starvation seems to reflect differences in control of nutritional or energy metabolism rather than differences in ability to synthesize ppGpp.
Construction of an Escherichia coli strain unable to synthesize putrescine, spermidine, or cadaverine: characterization of two genes controlling lysine decarboxylase
Authors: Tabor H , Hafner E W , Tabor C W
Published: January 2020

We have previously described a polyamine-deficient strain of Escherichia coli that contained deletions in speA (arginine decarboxylase), speB (agmatine ureohydrolase), speC (ornithine decarboxylase), and speD (adenosylmethionine decarboxylase). Although this strain completely lacked putrescine and spermidine, it was still able to grow at a slow rate indefinitely on amine-deficient media. However, these cells contained some cadaverine (1,5-diaminopentane). To rule out the possibility that the presence of cadaverine permitted the growth of this strain, we isolated a mutant (cadA) that is deficient in cadaverine biosynthesis, namely, a mutant lacking lysine decarboxylase, and transduced this cadA gene into the delta (speA-speB) delta speC delta D strain. The resultant strain had essentially no cadaverine but showed the same phenotypic characteristics as the parent. Thus, these results confirm our previous findings that the polyamines are not essential for the growth of E. coli or for the replication of bacteriophages T4 and T7. We have mapped the cadA gene at 92 min; the gene order is mel cadA groE ampA purA. A regulatory gene for lysine decarboxylase (cadR) was also obtained and mapped at 46 min; the gene order is his cdd cadR fpk gyrA.