bluuebunny/crossref_metadata_2025 huggingface dataset:
Unbalanced rRNA gene dosage and its effects on rRNA and ribosomal-protein synthesis
Authors: Siehnel R J , Morgan E A
Published: January 2020

The synthesis of rRNA was unbalanced by the introduction of plasmids containing rRNA operons with large internal deletions. Significant unbalanced synthesis was achieved only when the deletions affected both 16S and 23S RNA genes or when the deletions affected the 23S RNA gene alone. Although large imbalances in rRNA synthesis resulted from deletions affecting 16S and 23S RNA genes or only 23S RNA genes, excess 16S RNA and defective rRNA species were rapidly degraded. Large imbalances in the synthesis of regions of rRNA did not result in significantly unbalanced synthesis of ribosomal proteins. It therefore is probable that excess intact 16S RNA is degraded because ribosomal proteins are not available for packaging the RNA into ribosomes. Defective RNA species also may be degraded for this reason or because proper ribosome assembly is prevented by the defects in RNA structure. We propose two possible explanations for the finding that unbalanced overproduction of binding sites for feedback ribosomal protein does not result in significant unbalanced translational feedback depression of ribosomal protein mRNAs.
Glutathione-independent isomerization of maleylpyruvate by Bacillus megaterium and other gram-positive bacteria
Authors: Hagedorn S R , Bradley G , Chapman P J
Published: January 2020

Maleylpyruvate, the ring fission product of gentisic acid, was found to be isomerized to fumarylpyruvate without a requirement for glutathione by an enzyme activity found in cell extracts of m-hydroxybenzoate-grown Bacillus megaterium 410. The isomerization reaction was detected as a shift in the absorbance maximum from 330 nm, the maximum for maleylpyruvate, to 345 nm, the maximum for fumarylpyruvate, when assayed at pH 8.0. Ammonium sulfate precipitation and dialysis of B. megaterium cell extracts resolved the isomerase activity from low-molecular-weight compounds such as glutathione but did not eliminate the isomerase activity. Iodoacetate and p-chloromercuribenzoate were potent inhibitors of the isomerase from B. megaterium. However, N-ethylmaleimide and iodoacetamide did not significantly inhibit this activity. In addition, fumaric acid was demonstrated as a product of gentisate oxidation by dialyzed cell extracts of B. megaterium. Glutathione-independent maleylpyruvate isomerases with properties similar to the isomerase found in B. megaterium were also found in other genera of gram-positive organisms. Eleven different organisms representing the genera Bacillus, Arthrobacter, Corynebacterium, Nocardia, and Rhodococcus were all found to possess this novel type of glutathione-independent maleylpyruvate isomerase.
Purification and characterization of selenocysteine beta-lyase from Citrobacter freundii
Authors: Chocat P , Esaki N , Tanizawa K , Nakamura K , Tanaka H , Soda K
Published: January 2020

The purification and characterization of bacterial selenocysteine beta-lyase, an enzyme which specifically catalyzes the cleavage of L-selenocysteine to L-alanine and Se0, are presented. The enzyme, purified to near homogeneity from Citrobacter freundii, is monomeric with a molecular weight of ca. 64,000 and contains 1 mol of pyridoxal 5'-phosphate as a cofactor per mol of enzyme. L-Selenocysteine is the sole substrate (Km, 0.95 mM). L-Cysteine is a competitive inhibitor of the enzyme (Ki, 0.65 mM). The enzyme also catalyzes the alpha, beta elimination of beta-chloro-L-alanine to form NH3, pyruvate, and Cl- and is irreversibly inactivated during the reaction. The physicochemical properties, e.g., amino acid composition and subunit structure, of the bacterial enzyme are fairly different from those of the pig liver enzyme (Esaki et al., J. Biol. Chem. 257:4386-4391, 1982). However, the catalytic properties of both enzymes, e.g., substrate specificity and inactivation by the substrate or a mechanism-based inactivator, beta-chloro-L-alanine, are very similar.
Sequence of the Escherichia coli pheST operon and identification of the himA gene
Authors: Mechulam Y , Fayat G , Blanquet S
Published: January 2020

The complete nucleotide sequence of the Escherichia coli pheST operon coding for the two subunits of phenylalanyl-tRNA synthetase (an alpha 2 beta 2-type enzyme) has been determined. Another open reading frame (prp) was revealed downstream from pheT which was identified as himA, the gene for the alpha subunit of the integration host factor.
Genetic recombination of bacterial plasmid DNA: effect of RecF pathway mutations on plasmid recombination in Escherichia coli
Authors: Kolodner R , Fishel R A , Howard M
Published: January 2020

Tn5 insertion mutations in the recN gene, and in what appears to be a new RecF pathway gene designated recO and mapping at approximately 55.4 min on the standard genetic map, were isolated by screening Tn5 insertion mutations that cotransduced with tyrA. The recO1504::Tn5 mutation decreased the frequency of recombination during Hfr-mediated crosses and increased the susceptibility to killing by UV irradiation and mitomycin C when present in a recB recC sbcB background, but only increased the sensitivity to killing by UV irradiation when present in an otherwise Rec+ background. The effects of these and other RecF pathway mutations on plasmid recombination were tested. Mutations in the recJ, recO, and ssb genes, when present in otherwise Rec+ E. coli strains, decreased the frequency of plasmid recombination, whereas the lexA3, recAo281, recN, and ruv mutations had no effect on plasmid recombination. Tn5 insertion mutations in the lexA gene increased the frequency of plasmid recombination. These data indicate that plasmid recombination events in wild-type Escherichia coli strains are catalyzed by a recombination pathway that is related to the RecF recombination pathway and that some component of this pathway besides the recA gene product is regulated by the lexA gene product.
F factor inhibition of conjugal transfer of broad-host-range plasmid RP4: requirement for the protein product of pif operon regulatory gene pifC
Authors: Miller J F , Lanka E , Malamy M H
Published: January 2020

By the use of deletions, point mutations, and gene fusions, we show that the protein product of the F factor pifC gene is responsible for F factor inhibition of plasmid RP4 conjugal transfer. Deletion analysis of pif sequences carried by pSC101-F chimeric plasmids demonstrated that removal of all or part of the pifC coding sequence greatly decreased or abolished the ability of these plasmids to inhibit RP4 transfer. Amber mutations in the pifC gene eliminated inhibition in an Su- host strain but not in and Su+ (supF) host. Plasmids carrying nonpolar pifC mutations did not decrease the efficiency of RP4 transfer when present in trans. Whereas pifC+ plasmids inhibited RP4 transfer, the presence of RP4 in the same cell as F' lac increased F'lac Pif activity approximately 1,000-fold. This effect most likely resulted from the binding of the pifC product to RP4 DNA and concomitant derepression of the F factor pif operon. PifC inhibited trans mobilization of pMS204, a nonconjugative plasmid carrying the RP4 oriT locus, by the RP1 derivative pUB307. pMS204 had no trans effect on pif operon expression, whereas pUB307 increased F'lac Pif expression, as did RP4. Our results suggest that the pifC product inhibits expression of one or more RP4 genes, the products of which are required for conjugal transfer of RP4 and are required in trans for mobilization of nonconjugal RP4 oriT containing plasmids.
Effect of changes in the osmolarity of the growth medium on Vibrio cholerae cells
Authors: Lohia A , Majumdar S , Chatterjee A N , Das J
Published: January 2020

The rate and extent of lysis of Vibrio cholerae cells under nongrowing conditions were dependent on the osmolarity of the growth medium. Gross alterations in cellular morphology were observed when V. cholerae cells were grown in media of high and low osmolarity. The rate of lysis of V. cholerae cells under nongrowing conditions increased after treatment with chloramphenicol. Chloramphenicol-treated V. cholerae 569B cells showed formation of sphaeroplast-like bodies in medium of high osmolarity, but not in low osmolarity. Changes in the osmolarity of the growth medium also regulated the expression of the outer membrane proteins. This regulation was abolished if V. cholerae cells were grown in Pi-depleted medium. Analysis of the lytic behavior and composition of outer membrane proteins of an osmotically fragile mutant strain revealed a similar dependence on the osmolarity of the growth medium.
Effect of Calcofluor white and Congo red on fungal cell wall morphogenesis: in vivo activation of chitin polymerization
Authors: Roncero C , Durán A
Published: January 2020

Calcofluor White (a fluorochrome dye) affected the growth of Geotrichum lactis by causing lysis of cells at the hyphal tips. This effect was prevented by the presence of an osmotic stabilizer. The growth of Saccharomyces cerevisiae was also affected, and multicellular aggregates were formed because of incomplete separation of mother and daughter cells; fluorescence microscopy indicated the presence of abnormally thick septa. The formation of rudimentary wall material by G. lactis protoplasts was promoted by Calcofluor or Congo red (another dye). The rate of chitin synthesis in protoplasts and growing cells was enhanced by both dyes. In contrast, both dyes inhibited chitin and beta (1,3)-glucan synthases in isolated cell-free systems. Degradation rates for beta (1,3)-glucan or chitin were not affected significantly by the dyes. Wheat germ agglutinin also affected chitin synthesis in protoplasts.
Multiple controls exerted on in vivo expression of the pepN gene in Escherichia coli: studies with pepN-lacZ operon and protein fusion strains
Authors: Gharbi S , Belaich A , Murgier M , Lazdunski A
Published: January 2020

Three physiological conditions were shown to promote transcriptional regulation of pepN expression: phosphate limitation, the nature of the source of carbon and energy, and anaerobiosis. The transcriptional level of regulation can be deduced from the observation of these effects in strains carrying operon fusion pepN-lacZ. Mutations in the various genes phoB, phoM, phoR, crp, and fnr (oxrA) did not affect pepN expression.
Sodium and proton transport in Mycoplasma gallisepticum
Authors: Linker C , Wilson T H
Published: January 2020

When washed cells of Mycoplasma gallisepticum were incubated at 37 degrees C in 250 mM 22NaCl, the intracellular Na+ increased, and the K+ decreased. The addition of glucose to these Na+-loaded cells caused Na+ efflux and K+ uptake (both ions moving against concentration gradients). This effect of glucose was blocked by the ATPase inhibitor dicyclohexylcarbodiimide, which prevents the generation of a proton motive force in these cells. In additional experiments, Na+ extrusion was studied by diluting the 22Na+-loaded cells into Na+-free media and following the loss of 22Na+ from the cells. Glucose stimulated 22Na+ extrusion in such cells by a dicyclohexylcarbodiimide-sensitive mechanism. Proton movement was studied by measuring the pH gradient across the cell membrane with the 9-aminoacridine fluorescence technique. Glucose addition to cells preincubated with cations other than Na+ resulted in cell alkalinization (which was prevented by dicyclohexylcarbodiimide). This observation is consistent with the operation of a proton-extruding ATPase. When glucose was added to Na+-loaded cells and diluted into Na+-free media, intracellular acidification was observed, followed several minutes later by a dicyclohexylcarbodiimide-sensitive alkalinization process. The initial acidification was probably due to the operation of an Na+-H+ antiport, since Na+ exit was occurring simultaneously with H+ entry. When Na+-loaded cells were diluted into Na+-containing media, the subsequent addition of glucose resulted in a weak acidification, presumably due to H+ entry in exchange for Na+ (driven by the ATPase) plus a continuous passive influx of Na+. All of the data presented are consistent with the combined operation of an ATP-driven proton pump and an Na+ -H+ exchange reaction.
Repair of cis-platinum-DNA adducts by ABC excinuclease in vivo and in vitro
Authors: Husain I , Chaney S G , Sancar A
Published: January 2020

cis-Platinum compounds, which are used in cancer chemotherapy, are thought to exert their effect by damaging DNA. It is known that this damage is partially repaired in Escherichia coli. Using cis-Pt-treated pBR322 DNA as a probe, we investigated the role of nucleotide excision repair in the removal of Pt-DNA adducts. We found that the nucleotide excision pathway was the major mechanism for repairing Pt adducts in transforming plasmid DNA but that a recA-dependent pathway also contributed to plasmid survival. When cis-Pt-damaged pBR322 was treated with the purified nucleotide excision enzyme ABC excinuclease in vitro, a fraction of the adducts was removed by the enzyme; this removal resulted in a corresponding increase in transformation efficiency.
fipB and fipC: two bacterial loci required for morphogenesis of the filamentous bacteriophage f1
Authors: Lopez J , Webster R E
Published: January 2020

We describe the identification of two mutations in bacterial genes, designated as fipB and fipC, which resulted in temperature-sensitive morphogenesis of bacteriophage f1. These mutations mapped at separate loci but had to be present simultaneously to block f1 production at 41.5 degrees C. One mutation defined the locus fipB at 85.3 min on the Escherichia coli linkage map; the other defined the locus fipC, which mapped very close to rpsL at 73 min. Since these mutations did not appear to affect phage DNA replication, gene expression, or protein localization, they probably interfered with the its life cycle at the level of assembly. fipB mutants were partially deficient in adsorption of bacteriophage lambda, and fipB and fipC mutants leaked beta-lactamase into the medium, suggesting that the mutations affect outer-membrane structure or function.
Natural relationship between bacteroides and flavobacteria
Authors: Weisburg W G , Oyaizu Y , Oyaizu H , Woese C R
Published: January 2020

Comparisons among 16S rRNA sequences from various eubacteria reveal a natural relationship between the bacteroides (represented by the Bacteroides fragilis sequence) and a phylogenetic unit that comprises the flavobacteria, cytophagae, flexibacteria, and others (represented by the Flavobacterium heparinum sequence). Although the relationship is not a close one, it is, nevertheless, specific. rRNAs from these two organisms are not only closer to one another in overall sequence than they are to outgroup species (such as Bacillus subtilis, Escherichia coli, Desulfovibrio desulfuricans, and Agrobacterium tumefaciens), but they show common idiosyncrasies (i.e., derived characteristics) in both rRNA sequences and higher-order structures.
Electrophoretic and immunochemical analyses of the lipopolysaccharides from various strains of Aeromonas hydrophila
Authors: Dooley J S , Lallier R , Shaw D H , Trust T J
Published: January 2020

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the lipopolysaccharides isolated from strains of Aeromonas hydrophila which exhibit virulence for fish and which autoaggregate during growth in static broth culture. The lipopolysaccharides contained O-polysaccharide chains of homogeneous chain length. Two of the strains produced a surface protein array, and immunofluorescence and phage-binding studies revealed that a number of these O-polysaccharide chains of homogeneous length traversed the protein array and were exposed on the cell surface. Immunochemical analyses by immunoblotting, enzyme-linked immunosorbent assay, immunofluorescence, and immunoprecipitation with both polyclonal and monoclonal antibodies revealed the presence of three epitopes on the polysaccharide moiety of this homogenous-chain-length lipopolysaccharide morphotype. One epitope was species serogroup specific and reactive by immunoblotting. This epitope was not present on the heterogeneous-chain-length O polysaccharides of nonautoaggregating strains of A. hydrophila examined. The second epitope was conformation dependent and cross-reactive with an epitope on the homogenous-chain-length O polysaccharides of Aeromonas salmonicida lipopolysaccharide. The third epitope was recognized by a monoclonal antibody and appeared to involve that region of the A. hydrophila and A. salmonicida lipopolysaccharide molecules which contained the O-polysaccharide-core oligosaccharide glycosidic linkage.
Mechanism of bacteriophage conversion of lipase activity in Staphylococcus aureus
Authors: Lee C Y , Iandolo J J
Published: January 2020

Staphylococcus aureus PS54 harbors two temperate bacteriophages and manifests no lipase activity on egg yolk agar. Curing of one of the resident prophages (L54a) restores lipase activity. To study the mechanism of bacteriophage conversion, the prophage was cured, and the gene encoding lipase activity was cloned into pBR322 in Escherichia coli on a 2.9-kilobase DNA fragment of the chromosome. The fragment was subcloned into a shuttle vector and subsequently transformed into S. aureus and Bacillus subtilis. Lipase activity was expressed in all three genetic backgrounds. Transformation and transductional data indicated that conversion is due to insertional inactivation of the lipase gene. Hybridization analysis with probes made from converting-phage DNA and from the cloned fragment confirmed that the phage insertion site resides within the terminal 0.8 kilobase of the insert.
Cloning and expression of bacterial ice nucleation genes in Escherichia coli
Authors: Orser C , Staskawicz B J , Panopoulos N J , Dahlbeck D , Lindow S E
Published: January 2020

Epiphytic populations of Pseudomonas syringae and Erwinia herbicola are important sources of ice nuclei that incite frost damage in agricultural crop plants. We have cloned and characterized DNA segments carrying the genes (ice) responsible for the ice-nucleating ability of these bacteria. The ice region spanned 3.5 to 4.0 kilobases and was continuous over this region in P. syringae Cit7R1. The cloned fragments imparted ice-nucleating activity in Escherichia coli. Substantial increases in the nucleating activity of both E. coli and P. syringae were obtained by subcloning the DNA fragments on multicopy plasmid vectors. Southern blot analysis showed substantial homology between the ice regions of P. syringae and E. herbicola, although individual restriction sites within the ice regions differed between the two species.
recA is required in the induction of pectin lyase and carotovoricin in Erwinia carotovora subsp. carotovora
Authors: Zink R T , Engwall J K , McEvoy J L , Chatterjee A K
Published: January 2020

Pectin lyase (PNL) and the bacteriocin carotovoricin (CTV) were induced in Erwinia carotovora subsp. carotovora 71 by the DNA-damaging agents mitomycin C, nalidixic acid, and UV light. To determine whether the recA product was involved in the expression of these damage-inducible phenotypes, we cloned the E. carotovora subsp. carotovora recA+ gene, inactivated it by Tn5 insertion, and constructed an E. carotovora subsp. carotovora recA::Tn5 strain by gene replacement via homologous recombination. The RecA- strain was more sensitive to methyl methanesulfonate, nitroquinoline oxide, and UV light than its RecA+ parent. The recA mutation did not affect the production of pectate lyase, polygalacturonase, cellulase, and protease or the ability to cause soft rot of potato tubers. With this mutant, unlike with the RecA+ parent strain, PNL and CTV were not induced by mitomycin C or detected in potato tuber tissue. The RecA+ phenotype, including the inducibility of PNL and CTV, could, however, be restored in the mutant in trans by the recA+ gene from either E. carotovora subsp. carotovora or Escherichia coli. We conclude that, in E. carotovora subsp. carotovora, the recA product is required in the induction of PNL and CTV.
Bacteriophages in L form of Staphylococcus aureus
Authors: Schmid E N
Published: January 2020

Lysogenicity and phage typability of Staphylococcus aureus L-form cells are described. Spontaneously produced phages were found in thin sections of S. aureus L colonies. The dimensions of the tail and head resemble those of the morphological group BIII2 (T. Krzywy, I. Durlakowa, A. Kucharewcz-Krukowska, S. Krynski, and S. Slopek, Zentralbl. Bakteriol. Mikrobiol. Hyg. Abt. 1 Orig. Reihe A 250:287-295, 1981). Phages 3A and 3C of the international typing set lysed both the bacillary and L form of S. aureus.
Autoplaquing in Neisseria gonorrhoeae
Authors: Campbell L A , Short H B , Young F E , Clark V L
Published: January 2020

Irregularly shaped autoplaques were observed on a lawn of two different strains of Neisseria gonorrhoeae. Autoplaquing occurred on gonococcal genetic medium lacking arginine and was noninducible on complete gonococcal genetic medium. The cell density, incubation temperature, and agar base influenced autoplaquing. Single-colony suspensions varied in plaque morphology. We were unable to isolate a stable nonplaquing variant but separated strain RUN5287 into two plaquing phenotypes.
Accumulation of alpha,alpha-trehalose by Rhizobium bacteria and bacteroids
Authors: Streeter J G
Published: January 2020

Four strains of Rhizobium japonicum (61A76 and USDA 110, 123, and 138) were grown in eight different defined media. Regardless of the carbon or nitrogen source supplied, alpha, alpha-trehalose was the major carbohydrate (among mono- and disaccharides) accumulated by all four strains. After 7 to 9 days of growth, trehalose generally accounted for 90 to 100% of the mono- and disaccharides detected. None of the four strains would grow with trehalose as a carbon source, but the utilization of endogenous trehalose was demonstrated under carbon starvation conditions in water culture or when the carbon supply in a defined medium was exhausted. Under these conditions, a small amount of trehalose was lost from cells to the medium. In a survey of most of the serogroups of R. japonicum and several strains of other Rhizobium species, all strains tested were found to accumulate some trehalose. Trehalose concentrations varied widely; the highest concentration recorded was 41 micrograms/mg of dry weight. In all but six strains trehalose accounted for greater than 80% of the mono- and disaccharides in cells. Fast-growing strains of R. japonicum also accumulated small amounts trehalose. R. japonicum bacteroids also synthesized trehalose; the quantity in nodules varied in approximate correspondence to accumulation of trehalose by cultured bacteria. In young soybean nodules (29 days after planting), 45 to 80% of the trehalose was recovered in the cytosol. There were differences among R. japonicum strains in the retention of trehalose, and the proportion of trehalose retained by bacteroids increased with increasing plant age for all strains.
Regulation of Pseudomonas aeruginosa chemotaxis by the nitrogen source
Authors: Craven R , Montie T C
Published: January 2020

The regulation of amino acid chemotaxis by nitrogen was investigated in the gram-negative bacterium Pseudomonas aeruginosa. The quantitative capillary tube technique was used to measure chemotactic responses of bacteria to spatial gradients of amino acids and other attractants. Chemotaxis toward serine, arginine, and alpha-aminoisobutyrate was sharply dependent on the form in which nitrogen was presented to the bacteria. Bacteria grown on mineral salts-succinate with potassium nitrate gave responses to amino acids that were 2 to 3 times those of cells grown on ammonium sulfate and 10 to 20 times those of cells grown in mineral salts-succinate with Casamino Acids as the nitrogen source. A combination of ammonium sulfate and glutamate was as effective as Casamino Acids in depressing serine taxis. The threshold concentration for alpha-aminoisobutyrate taxis was consistently lower in nitrate-grown bacteria than in ammonia-grown bacteria. Responsiveness to sodium succinate, however, was not subject to regulation by nitrogen, and glucose chemotaxis was inhibited, rather than enhanced, in nitrate-grown bacteria. These results indicate that chemotaxis of P. aeruginosa toward amino acids is subject to regulation by nitrogen and that this regulation probably is expressed at the level of the chemoreceptors or transducers.
Characterization of the in vivo RNA product of the pOUT promoter of IS10R
Authors: Lee Y , Schmidt F J
Published: January 2020

We characterized a single RNA species (RNAout1) which was the major in vivo RNA made from pOUT of IS10R. RNAout1 was 70 nucleotides long; its 5' end corresponded exactly to the in vitro start of pOUT transcription. The concentration of RNAout1 was estimated at 5 to 10 molecules per cell containing the single-copy plasmid NR1. RNA sequences from pOUT of IS10L were detected at a much lower (less than one molecule per cell) steady-state concentration and may be preferentially degraded in vivo. We suggest that the low level of the IS10L transcript led to the inability of IS10L sequences to translationally inhibit Tn10 transposition.
Domains involved in osmoregulation of the ompF gene in Escherichia coli
Authors: Inokuchi K , Itoh M , Mizushima S
Published: January 2020

Expression of the ompF gene, which is under the control of the OmpR protein, is regulated by the osmolarity of the medium. To study the mechanism of osmoregulation, plasmids carrying two different types of chimeric genes were constructed. In one type, the coding region of the ompF gene was linked to the trp promoter (trpPO) preceding ompF, and in the other type the ompF upstream region, mostly composed of the region for regulation by OmpR and the promoter region, was linked to the lacZ gene by protein fusion. Expression of beta-galactosidase by the lacZ chimeric gene was OmpR dependent and osmoregulated as sensitively as that of the intact ompF gene. In the ompR20 background the direction of osmoregulation was opposite that of normal osmoregulation, as was the direction of osmoregulation of the intact ompF gene. Osmoregulation was also observed with trpPO-ompF chimeric genes. However, the regulation was not as sensitive to the osmolarity of the medium as was regulation of the intact ompF gene and was independent of OmpR. These results suggest that OmpR-dependent osmoregulation played a primary role in the osmoregulation of ompF expression and that ompR-independent osmoregulation most likely did not play a crucial role. Studies with a series of trpPO-ompF chimeric genes also suggest that the untranslated leader region, about 100 base pairs in length, between the transcription initiation site and the initiation codon was not required for osmoregulation.
Methanosarcina mutant unable to produce methane or assimilate carbon from acetate
Authors: Smith M R , Lequerica J L
Published: January 2020

Mutants of Methanosarcina barkeri 227 resistant to monofluoroacetate were isolated from monofluoroacetate-treated cultures. Mutant strain FAr9 was 100 times more resistant to monofluoroacetate than the wild-type strain and was deficient in carbon uptake and CH4 and CO2 production from methyl-labeled acetate. Methanol was assimilated at increased levels. Strain FAr9 was unable to shift from using methanol to using acetate for growth and exhibited increased sensitivity to growth inhibition by NaCN in methanol-containing complex medium. Unlike parent strain 227, acetate addition to methanol-containing media did not prevent NaCN inhibition. The specific activities of enzymes of exogenous acetate assimilation, CO dehydrogenase, and enzymes of the tricarboxylic acid cycle were similar for mutant and parent strain cell extracts. Mutation to monofluoroacetate resistance did not confer simultaneous resistance to 2-bromoethanesulfonate or pyruvate or alter propionate uptake. We conclude that strain FAr9 is either an acetate permeability mutant or is defective in an activation step required for the catabolism and anabolism of acetate.
Isolation of insertion, deletion, and nonsense mutations of the uracil-DNA glycosylase (ung) gene of Escherichia coli K-12
Authors: Duncan B K
Published: January 2020

Two uracil-DNA glycosylase (ung) mutation selection procedures based upon the ability of uracil glycosylase to degrade the chromosomes of organisms containing uracil-DNA were devised to obtain a collection of well-defined ung alleles. In an enrichment procedure, lysogens were selected from Escherichia coli cultures infected with lambda pKanr phage containing uracil in their DNA. (These uracil-DNA phage were prepared by growth on host cells deficient in both dUTPase and uracil-DNA glycosylase.) The lysogenic Kanr population was enriched for uracil glycosylase-deficient mutants by a factor of 10(4). In a phage suicide selection procedure, lambda pung+ phage were unable to form plaques on dut ung cells containing uracil-DNA in their chromosomes, and all of the progeny were lambda pung-. Deletion, insertion (ung::Mu and ung::Tn10), nonsense, and missense mutants were isolated by using these procedures. Extracts of three insertion mutants contained no detectable enzyme activity. All of the other mutant isolates had less than 1% of the normal uracil glycosylase specific activity. The previously studied ung-1 allele, which was derived by N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis, produced about 0.02% of the normal amount of uracil glycosylase activity. No significant phenotypic differences between ung-1 and ung::Tn10 alleles were observed. Variations of the lysogen selection procedure may be helpful for isolating other DNA glycosylase mutations in E. coli and other organisms.
Evidence for transcription antitermination control of tryptophanase operon expression in Escherichia coli K-12
Authors: Stewart V , Yanofsky C
Published: January 2020

Tryptophanase, encoded by the gene tnaA, is a catabolic enzyme distinct from the enzymes of tryptophan biosynthesis. Tryptophanase synthesis is induced by tryptophan and is subject to catabolite repression. We studied the mechanism of tna operon induction. Mutants with altered rho factor were partially constitutive for tna expression, implicating rho-dependent transcription termination in the control of tna expression. Measurements of mRNA synthesis from the transcribed leader region preceeding the tna operon suggested that the tna promoter was constitutive and that in the absence of inducer, transcription terminated in the leader region. Upon induction, this transcription termination was relieved. Cis-acting constitutive mutants had genetic alterations in the tna leader region. These lesions defined a site that is homologous to the bacteriophage lambda boxA sequence, which is thought to play a role in antitermination control of lambda lytic gene expression. We propose that tna expression is subject to transcription antitermination control. We hypothesize that a tryptophan-activated antiterminator protein mediates induction by suppressing the rho-dependent termination sites in the leader region, thus allowing transcription to proceed into the tna operon structural gene region.
Diauxic growth in Azotobacter vinelandii
Authors: George S E , Costenbader C J , Melton T
Published: January 2020

Azotobacter vinelandii exhibited diauxie when grown in a medium containing both acetate and glucose as carbon sources. Acetate was used as the primary carbon source during the acetate-glucose diauxie. Uptake of acetate was constitutively expressed during both diauxic phases of growth. Induction of the glucose uptake system was inhibited in the presence of acetate. Acetate was also the preferred growth substrate for A. vinelandii grown in a medium containing either fructose, maltose, xylitol, or mannitol. The tricarboxylic acid cycle intermediates citrate, isocitrate, and 2-oxoglutarate inhibited glucose utilization in cells grown in glucose medium containing these substrates, and diauxic growth was observed under these growth conditions. Temporal expression of isocitrate-lyase, ATPase, and nitrogenase was exhibited during acetate-glucose diauxie.
IS186: an Escherichia coli insertion element isolated from a cDNA library
Authors: Kothary R K , Jones D , Candido E P
Published: January 2020

Escherichia coli IS186 was isolated from cDNA libraries made from rainbow trout RNA and maintained in E. coli RR1. The element was 1,347 base pairs in length, had a perfect inverted repeat of 25 base pairs, and had an open reading frame of 375 amino acids. The hypothetical protein sequence of IS186 had limited homology to the E. coli IS4 hypothetical protein I sequence. There were three copies of IS186 in E. coli RR1.
Isolation and characterization of Paracoccus denitrificans mutants with defects in the metabolism of one-carbon compounds
Authors: Harms N , de Vries G E , Maurer K , Veltkamp E , Stouthamer A H
Published: January 2020

Mutants deficient in the metabolism of one-carbon compounds have been obtained by treating Paracoccus denitrificans with the mutagen N-methyl-N'-nitro-N-nitrosoguanidine. Mutants were selected without enrichment procedures by newly developed plate screening tests. The obtained mutants were characterized by their growth responses, cytochrome composition, enzyme activities, and immunogenic reaction with antisera against methanol dehydrogenase. By these criteria five mutant classes could be distinguished. Class I mutants are involved in the expression of methanol dehydrogenase. Three mutants of this class have a defect in the structural gene. A double mutant was found with defects in the expression of both methanol dehydrogenase and hydrogenase. Class II mutants have a defect in a regulatory gene involved in the regulation of both methanol dehydrogenase and methylamine dehydrogenase. Class III mutants are deficient in formaldehyde metabolism. A defect may exist in the expression of a second non-NAD-linked formaldehyde dehydrogenase which was postulated to be involved in C1 metabolism. Class IV mutants are deficient in cytochrome c. Mutants of class V have a defect in synthesis of the molybdenum cofactor essential for the function of formate dehydrogenase.
Bacteriophage SPO2-mediated plasmid transduction in transpositional mutagenesis within the genus Bacillus
Authors: Kopec L K , Yasbin R E , Marrero R
Published: January 2020

A single copy of the Streptococcus faecalis transposon Tn917, located in the Bacillus subtilis chromosome, was able to transpose onto the SPO2 cos plasmid pPL1017, which codes for chloramphenicol resistance and contains the bacteriophage phi 105 immunity region. Selection for pPL1017::Tn917 chimeras was performed by SPO2-mediated plasmid transduction of transposon-borne resistance to macrolide-lincosamide-streptogramin B antibiotics (MLSr). The transposition of Tn917 onto plasmid pPL1017 occurred with a frequency of 10(-5) and was dependent on the presence of a subinhibitory dose of erythromycin. Twelve chimeras were subjected to genetic and physical analyses. Two Cams transductants harbored plasmids whose chloramphenicol acetyltransferase genes had been insertionally inactivated by Tn917. Several transpositions in the vicinity of the phi 105 immunity region were detected. However, all of the 300 MLSr, Camr transductants screened were immune to phi 105 infectious activity. One pPL1017::Tn917 chimera, pLK200, was transferred by SPO2 plasmid transduction into the Bacillus amyloliquefaciens prototrophic strain DSM7. Plasmid pLK200 was effective in the mutagenesis of the DSM7 chromosome and yielded auxotrophs at a frequency of 0.5 to 5.3%. Generation of auxotrophs was also dependent on the presence of a subinhibitory dose of erythromycin. Forty-four auxotrophs representing at least nine amino acid requirements were recovered.
Use of resistant mutants to study the interaction of triton X-100 with Staphylococcus aureus
Authors: Raychaudhuri D , Chatterjee A N
Published: January 2020

Staphylococcus aureus mutants resistant to the nonionic detergent Triton X-100, isolated from the wild-type strain H and the autolysin-deficient strain RUS3, could grow and divide in broth containing 5% (vol/vol) Triton X-100, while growth of the parental strains was markedly inhibited above the critical micellar concentration (0.02%) of the detergent. Growth-inhibitory concentrations of Triton X-100 killed wild-type cells without demonstrable cellular lysis. Triton X-100 stimulated autolysin activity of S. aureus cells under nongrowing conditions, and this lytic response was markedly reduced in energy-poisoned cells. In contrast, the detergent had no effect on the activity of autolysins in cell-free systems, and growth in the presence of Triton X-100 did not alter either the cellular autolysin activity or the susceptibility of cell walls to exogenous lytic enzymes. Treatment with either Triton X-100 or penicillin G in the growth medium stimulated release of predominantly acylated intracellular lipoteichoic acid and sensitized staphylococci to Triton X-100-induced autolysis. There was no significant difference in the cell wall and membrane compositions or Triton X-100 binding between the parental strains and the resistant mutants. The resistant mutant TXR1, derived from S. aureus H, had a higher level of L-alpha-glycerophosphate dehydrogenase activity, and its oxygen uptake was more resistant to inhibition by a submicellar concentration (0.008%) of Triton X-100. Growth in the presence of subinhibitory concentrations of Triton X-100 rendered S. aureus H cells phenotypically resistant to the detergent and greatly stimulated the level of oxygen uptake. Membranes isolated from such cells exhibited enhanced activity of the respiratory enzymes succinic dehydrogenase and L-alpha-glycerophosphate dehydrogenase.
lac Up-promoter mutants with increased homology to the consensus promoter sequence
Authors: Mandecki W , Goldman R A , Powell B S , Caruthers M H
Published: January 2020

Four lac promoter mutants were constructed. The mutations increased the homology between the lac promoter and the consensus promoter sequences by introducing the consensus -10 and -35 regions and the consensus spacing of 17 residues between these two regions. The promoter mutants were cloned into a pBR322-derivatized vector upstream from the lacZ gene, and levels of beta-galactosidase were an indication of promoter activity. All mutants exhibited higher activity than did the wild-type promoter.
Excretion of unassembled hook-associated proteins by Salmonella typhimurium
Authors: Homma M , Iino T
Published: January 2020

Hook-associated proteins (HAPs) were excreted into the culture medium of the Fla+ strain as well as into the growth medium of the filamentless mutants of Salmonella typhimurium. This indicates that the bacteria synthesize HAPs excessively, beyond the amount required for construction of flagella. The extra HAPs are shed into the culture medium after a definite amount of each HAP has been assembled into the flagellar structure.
PRODUCTION OF VIBRIO EL TOR TOXIN IN A LIQUID MEDIUM
Authors: Watanabe Yoshikazu , Felsenfeld Oscar
Published: January 2020

Watanabe, Yoshikazu(U. S. Army Medical Center, Japan),and Oscar Felsenfeld. Production of vibrio El Tor toxin in a liquid medium. J. Bacteriol.82:43–47. 1961.—High toxin titers (as measured by mouth lethality and hemolytic activity) were obtained from culture supernatants of strains of El Tor vibrio isolated from patients in Indonesia. Cultivation of the organism in vitro revealed that the concentration of ferrous ion in the medium was critical for the production of maximal yields of toxin.

Toxins produced under appropriate conditions were antigenic in rabbits and produced antibodies capable of neutralizing both the hemolytic and lethal actions in multiple proportion. Suggestive evidence has been obtained that the hemolytic and lethal toxins may be separate molecules.
FATE OF PARTIALLY PURIFIED C14-LABELED TOXIN OFCLOSTRIDIUM PERFRINGENS
Authors: Ellner Paul D.
Published: January 2020

Ellner, PaulD. (University of Florida, Gainesville). Fate of partially purified C14-labeled toxin ofClostridium perfringens. J. Bacteriol.82:275–283. 1961.—A study was made of the fate ofClostridium perfringenstoxin in susceptible animals. Labeled toxin was prepared by growingC. perfringenstype A in a complex casein hydrolyzate medium containing a tryptic hydrolyzate of C14-labeled algal protein. The toxin was purified about 30-fold (in terms of lecithinase activity) by ammonium sulfate and acetone precipitations. The tagged toxin was injected intravenously into mice and rabbits, and the disappearance from the blood stream, deposition in organs and appearance in urine and expired air determined by measurement of radioactivity. Experimental data showed that toxin disappears rapidly from the bloodstream following intravenous injection, with radioactivity appearing in the urine and expired air shortly thereafter (10 to 20 min). The organs primarily responsible for the uptake of toxin from the blood are the liver (72%), lungs (15%), kidney (8%), and spleen (5%). The toxin is not bound to skeletal muscle. Fractionation of the liver into subcellular particles by centrifugation showed the radioactivity to be concentrated in the mitochondrial fraction. These experiments indicate that the toxin is rapidly removed from the circulating blood, is metabolized, and breakdown products excreted in the urine, with 1 carbon fragments eliminated as CO2in the expired air.
PATHWAY FOR THE DISSIMILATION OF ITACONIC AND MESACONIC ACIDS
Authors: Brightman Vernon , Martin William R.
Published: January 2020

Brightman, Vernon
(The University of Chicago, Chicago),
and William R. Martin
. Pathway for the dissimilation of itaconic and mesaconic acids. J. Bacteriol.
82:
376–382. 1961.—Studies on the oxidation of itaconic and mesaconic acids by a
Pseudomonas
sp., adapted to utilize either of these acids as a sole carbon source, have provided evidence for a pathway converting both itaconate and mesaconate to succinate. A metabolic interconversion of itaconate, mesaconate, and citramalate has also been demonstrated by whole cell and cell-free enzyme studies.

Succinate derived from methylene-labeled itaconate was found to be labeled in the inside carbon atoms, a fact which indicates that the branched chain compound was converted into a straight chain molecule by a shift of the methylene carbon (C-5) from the side chain of itaconate to a position between C-2 and C-3 in an, as yet, unknown straight chain intermediate prior to its conversion to succinate.
FACTORS AFFECTING ASPARTASE ACTIVITY
Authors: Depue Robert H. , Moat Albert G.
Published: January 2020

Depue, Robert
H. (Hahnemann Medical College, Philadelphia),
and Albert G. Moat
. Factors affecting aspartase activity. J. Bacteriol.
82:
383–386. 1961.—Cells of
Escherichia coli
grown in a glucose-mineral salts medium contain about one-fifth the amount of aspartase activity observed in cells grown in a yeast extract-peptone medium. The aspartase activity of the cells grown in glucose-salts medium would appear to be too low to provide a mechanism for synthesis of amino groups. Aspartase was purified approximately eightfold by ammonium sulfate precipitation and column chromatography of cell-free extracts. The purified preparation was specific for
l
-aspartic acid and contained no fumarase activity. A divalent metal ion requirement was demonstrated, this requirement being satisfied by cobaltous or manganous ions. The enzyme activity was found to be dependent upon free sulfhydryl groups. Biotin did not appear to be directly involved in the aspartase reaction since high concentrations of avidin did not alter the reaction rate. The Michaelis constant for aspartase with aspartic acid as substrate was determined to be 0.033
m
.
METABOLIC CHARACTERIZATION OF THE GENUS
BRUCELLA
II
Authors: Meyer Margaret E. , Cameron H. S.
Published: January 2020

Meyer, Margaret
E. (University of California, Davis),
and H. S. Cameron
. Metabolic characterization of the genus
Brucella
. II. Oxidative metabolic patterns of the described biotypes. J. Bacteriol.
82:
396–400. 1961.—The oxidative metabolism of the biotypes of
Brucella abortus
and
Brucella suis
was compared to that of typical strains of these species. It was found that the biotypes of
B. abortus
display a metabolic pattern identical to that of
B. abortus
type I, irrespective of their differences in susceptibility to the bacteriostatic action of basic fuchsin and thionin.

It was also found that a characteristic feature which distinguishes all of the biotypes of
B. suis
from the other species was their ability to oxidize
l
-arginine,
dl
-citrulline, and
dl
-ornithine. The three biotypes of
B. suis
can be differentiated from each other by discrete differences in their utilization of
l
-lysine and
l
-glutamic acid.

Data on the oxidative rates obtained on eight amino acid and four carbohydrate substrates by 47 strains of the described biotypes are presented in support of these conclusions.
CITRIC ACID FERMENTATION OF SUGARS PURIFIED WITH CHELATING RESIN
Authors: Noguchi Yuichi , Johnson Marvin J.
Published: January 2020

Noguchi, Yuichi
(University of Wisconsin, Madison),
and Marvin J. Johnson
. Citric acid fermentation of sugars purified with chelating resin. J. Bacteriol.
82:
538–541. 1961.—A new, independently isolated strain of
Aspergillus niger
capable of giving high yields of citric acid in submerged culture was found to show the same behavior toward iron, zinc, and manganese as a previously studied strain. Citric acid accumulation did not occur in the presence of manganese. Best citric acid production was obtained in the presence of limited amounts of iron and zinc. Use of a chelating ion exchange resin was found to be an excellent method of removing polyvalent metals from sugars, either for analytical purposes or for fermentation. Commercial glucose, after resin treatment, gave citric acid yields of more than 80% when supplemented with iron and zinc. Unpurified glucose was converted to citric acid in good yields by mycelium grown on resin-treated glucose.
ROLE OF POTASSIUM IN THE OXIDATIVE METABOLISM OF
MICROCOCCUS SODONENSIS
Authors: Perry Jerome J. , Evans James B.
Published: January 2020

Perry, Jerome
J. (The University of Chicago, Chicago, Ill.),
and James B. Evans
. Role of potassium in the oxidative metabolism of
Micrococcus sodonensis
. J. Bacteriol.
82:
551–555. 1961.—An absolute potassium requirement has been established for the growth of
Micrococcus sodonensis
with lactate or pyruvate as substrate. Potassium at 0.67 × 10
−2
m
concentration was necessary for maximal growth. Resting cell and cell-free preparations from cells grown on minimal levels of potassium were stimulated by potassium but, due to residual or bound cation, did not show an absolute requirement. Rubidium and cesium replaced potassium in these cells although cesium is much less effective.
INFLUENCE OF A BACTERIAL CELL EXTRACT UPON THE MORPHOGENESIS OFARTHROBACTER UREAFACIENS
Authors: Blankenship L. C. , Doetsch R. N.
Published: January 2020

Blankenship, L. C. (University of Maryland, College Park)and R. N. Doetsch. Influence of a bacterial cell extract upon the morphogenesis ofArthrobacter ureafaciens. J. Bacteriol.82:882–888. 1961—The effect of a bacterial extract on alleviating the abnormal morphological appearance ofArthrobacter ureafacienswhen cultivated in Thiotone broth is described. Physical and chemical analyses of this extract revealed that the mineral components therein were largely responsible for the observed effects. Similar results were obtained using ashed Trypticase soy broth, known mineral mixtures, or magnesium ions. The role played by these materials in governing the morphology ofA. ureafaciensis discussed.