bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of a peptide transport-deficient mutant of yeast
Authors: Marder R , Rose B , Becker J M , Naider F
Published: January 2020

A peptide transport mutant of a leucine-lysine auxotroph of Saccharomyces cerevisiae (strain Z1-2D) was isolated on the basis of its resistance to L-ethionyl-L-alanine. The mutant, designated Z1-2D Etar, did not utilize di- and tripeptides containing leucine or lysine although it contained peptidases which released the required amino acids from these substrates. S. cerevisiae Z1-2D Etar did not accumulate radioactivity from [14C]glycyl-L-leucine under conditions identical to those in which the parent took up the label from this dipeptide. These results indicate that the mutant lacks the cellular mechanism to transport peptides to the site of the peptidase activity and that di- and tripeptides share a common mode of entry into yeast.
Expression of
Escherichia coli
K-12 Arginine Genes in
Pseudomonas fluorescens
Authors: Mergeay M. , Boyen A. , Legrain C. , Glansdorff N.
Published: January 2020

Escherichia coli argE
and
argH
gene products were detected in
Pseudomonas fluorescens argH122
carrying the
E. coli
F110 plasmid.
Effects of altered rho gene product on the expression of the Escherichia coli histidine operon
Authors: Lawther R P , Hatfield G W
Published: January 2020

An altered rho gene product affects expression of the his operon of Escherichia coli. The effect is greater for the operator proximal portion of the his operon than for the operator distal portion. This "rho effect" appears to be independent of the site of action of hisT-altered histidyl-tRNA.
Differential effect of hydroxyurea on the replication of plasmid and chromosomal DNA in Bacillus subtilis
Authors: Shivakumar A G , Dubnau D
Published: January 2020

The replication in Bacillus subtilis of the staphylococcal R plasmids pE194, pBD15, pUB110, pSA0501, and pSA2100 has been studied in the presence of hydroxyurea. In all cases, an enrichment for covalently closed circular DNA compared with chromosomal DNA was observed. In this respect, hydroxyurea mimics the effect previously observed with pUB110, using strains carrying the conditional mutation dnaA13. This mutation has been reported to affect ribonucleotide reductase (G. W. Bazill and D. Karamata, Mol. Gen. Genet. 117:19-29, 1972). An explanation for these effects is offered, together with some supporting evidence.
Mechanism of autoenergized transport and nature of energy coupling for D-lactate in Escherichia coli
Authors: Kang S Y
Published: January 2020

To fully energize the active transport systems of Escherichia coli, it is common practice to preincubate the cells for 10 min with 10 or 20 mM concentration of a compound that can serve as an energy source. This paper shows that the active accumulation of D-lactate can be achieved within 1 min with only 50 micron D-lactate serving as an energy source for its own uptake in starved cells (autoenergization). The cells were strain DL54 which had been induced by growth in the presence of D-lactate. Uninduced cells were not able to show autoenergized D-lactate uptake under these conditions. The induced cells were also able to transport proline in the presence of 100 micron D-lactate as sole energy source. The D-lactate-dependent dehydrogenase activity in inverted French press vesicles was comparable for the induced and uninduced cells. The same was true for respiration of whole cells in the presence of 20 mM D-lactate. However, the Vmax for D-lactate transport of induced cells was six times higher than that of uninduced cells. It appears that a sufficient number of high-affinity carrier molecules in the cytoplasmic membrane are necessary for the autoenergized transport of D-lactate. A similar conclusion was reached for the autoenergized uptake of glycerol-3-phosphate by Escherichia coli strain 7. The active transport of D-lactate is driven by the protonmotive force.
Relationship of Glycolytic Intermediates, Glycolytic Enzymes, and Ammonia to Glycogen Metabolism During Sporulation in the Yeast
Saccharomyces cerevisiae
Authors: Fonzi William A. , Shanley Mark , Opheim Dennis J.
Published: January 2020

To identify the factors which control glycogen synthesis in
Saccharomyces cerevisiae
, we have studied the regulation of glycogen metabolism during sporulation, since in vivo glycogen has been reported to undergo significant changes in concentration during this process. We examined the concentration of a number of key glycolytic intermediates and enzymes in strains that sporulate at different rates and those that are deficient in sporulation. There were no significant changes found in the adenylate energy charge or cyclic AMP levels throughout sporulation. Although significant alterations occurred in the levels of glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, and ATP during sporulation, only the fourfold increase in fructose-1,6-bisphosphate appeared to correlate with glycogen synthesis in all of the strains examined. Only limited changes occurred in the level of a number of glycolytic and gluconeogenic enzymes which were examined during this process. Intracellular glucose content underwent a dramatic 30- to 40-fold increase in sporulating cells. Comparison of strains with different rates of sporulation demonstrated that this increase in glucose content coincides with the time of glycogen degradation in each strain. Both the increase in glucose content and the degradation of accumulated glycogen were not observed in nonsporulating α/α strains, or in cells incubated in NH
4
+
supplemented sporulation medium. Although glucose appears to be the direct product of glycogen degradation, a 10-fold increase in a nonspecific alkaline phosphatase occurs at this time, which may be degrading phosphorylated sugars to glucose. All of the strains examined released extracellular glucose while suspended in acetate sporulation medium. It is concluded that most of the changes in the glycolytic pathway that occur during sporulation, with the exception of glycogen degradation and the concomitant increase in intracellular glucose pools, are a response to the transfer to sporulation medium and are independent of sporulation-specific processes. Inhibition of sporulation with ammonium ions resulted in a different pattern of change in all of the glycolytic intermediates examined, including a twofold increase in cyclic AMP levels. Ammonia did not interfere with glycogen synthesis, but prevented sporulation-specific glycogen degradation. The levels of the glycolytic enzymes examined were not affected by ammonia.
Isolation and characterization of an endogenous inhibitor of protein synthesis in Escherichia coli K-12
Authors: Clark V L
Published: January 2020

A low-molecular-weight factor was isolated from cell extracts of Escherichia coli K-12. The concentration of the factor in cells was dependent upon nutritional conditions, the concentration being higher in faster growing cells. Treatment of cells with colicin K caused an increase in concentration of the factor. The factor inhibited protein synthesis in E. coli. This inhibition was reversible, apparently because of metabolism of the factor. The inhibition of synthesis of beta-galactosidase lasted longer than the inhibition of protein synthesis; cyclic AMP eliminated this difference. The factor inhibited the synthesis of beta-galactosidase from preformed lac mRNA, indicating an inhibition of translation. Kinetic studies of the onset of inhibition of beta-galactosidase synthesis by the factor suggested that the factor may inhibit protein synthesis at the initiation of translation.
Phosphoglucose isomerase mutant of Rhizobium meliloti
Authors: Arias A , Cerveńansky C , Gardiol A , Martínez-Drets G
Published: January 2020

A mutant strain of complex phenotype was selected in Rhizobium meliloti after nitrosoguanidine mutagenesis. It failed to grow on mannitol, sorbitol, fructose, mannose, ribose, arabitol, or xylose, but grew on glucose, maltose, gluconate, L-arabinose, and many other carbohydrates. Assay showed the enzyme lesion to be in phosphoglucose isomerase (pgi), and revertants, which were of normal growth phenotype, contained the enzyme again. Nonpermissive substrates such as fructose and xylose prevented growth on permissive ones such as L-arabinose, and in such situations there was high accumulation of fructose 6-phosphate. The mutant strain had about 20% as much exopolysaccharide as the parent. Nitrogen fixation by whole plants was low and delayed when the mutant strain was the inoculant.
Crown gall teratoma formation is plasmid and plant controlled
Authors: Gresshoff P M , Skotnicki M L , Rolfe B G
Published: January 2020

Experiments using different species of the plant Nicotiana and strains of the bacterium Agrobacterium tumefaciens showed that teratoma formation from crown galls was dependent on the combination of bacterial Ti plasmid and host plant used.
Lipid and lipopolysaccharide composition of Escherichia coli surface-altered mutants selected for resistance to levallorphan, tetracaine, and polymyxin
Authors: Dame J B , Shapiro B M
Published: January 2020

Certain mutants of Escherichia coli with an altered permeability barrier have an essentially normal lipopolysaccharide, fatty acid, and phospholipid content, with a slight increase in the membrane protein:lipid ratio. The phospholipid metabolism of the lev and tec strains shows an abnormal response to growth in the selective agents levallorphan and tetracaine, respectively.
Synthesis of ribosomal proteins during the cell cycle of the yeast Saccharomyces cerevisiae
Authors: Elliott S G , Warner J R , McLaughlin C S
Published: January 2020

Centrifugal elutriation was used to separate yeast cells by their cell cycle position. The rate of synthesis of ribosomal proteins showed a constant exponential increase through the cell cycle.
Lipoprotein synthesis in Escherichia coli spheroplasts: accumulation of lipoprotein in cytoplasmic membrane
Authors: Kanazawa H , Wu H C
Published: January 2020

Synthesis of cell envelope proteins was studied in ethylenediaminetetraacetic acid-lysozyme spheroplasts of Escherichia coli ML30. The rate of incorporation of [3H]arginine into proteins in spheroplasts was about 30% of that of intact cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins synthesized in spheroplasts revealed the preferential synthesis of five polypeptides, one of which has been identified as the free form of murein lipoprotein. Lipoprotein synthesized in spheroplasts was found to be of same molecular size as that of mature lipoprotein. No prolipoprotein was observed even with a short pulse-labeling with [3H]arginine. On the other hand, significant accumulation of newly synthesized lipoprotein in the cytoplasmic membrane fraction of spheroplasts was observed. These results suggest that the processing of prolipoprotein occurs in the cytoplasmic membrane fraction of the cell envelope.
Control of aldehyde synthesis in the luminous bacterium Beneckea harveyi
Authors: Ulitzur S , Hastings J W
Published: January 2020

Some of the Beneckea harveyi dim aldehyde mutants, all of which emit light upon addition of exogenous long-chain aldehyde, also emit light when myristic acid is added. Analysis of these myristic acid-responsive mutants indicates that they are blocked before fatty acid formation, whereas another class of mutants, which respond only to aldehyde, appear to be defective in the enzyme(s) involved in the conversion of acid to aldehyde. Evidence is presented that this activity, designated myristic acid reductase, is coinduced with luciferase and is involved in the recycling of acid produced in the luciferase reaction, with specificity for the C14 compounds.
Synthesis of guanosine tetra- and pentaphosphates by the obligately anaerobic bacterium Bacteroides thetaiotaomicron in response to molecular oxygen
Authors: Glass T L , Holmes W M , Hylemon P B , Stellwag E J
Published: January 2020

Guanosine 5'-diphosphate 3'-diphosphate (ppGpp) and guanosine 5'-triphosphate 3'-diphosphate (pppGpp) were detected in formic acid extracts of air-exposed culutres of Bacteroides thetaiotaomicron. The identification of ppGpp and pppGpp in B. thetaiotaomicron was based on the following results: (i) cochromatography of 32P-labeled hyperphosphorylated nucleotides in two different two-dimensional solvent systems with authentic ppGpp and pppGpp; (ii) incorporation of [3H]guanosine into the putative ppGpp and pppGpp; (iii) alkaline lability; and (iv) resistance, to periodate oxidation. There was a marked increase in the concentration of ppGpp and pppGpp after shift from anaerobic to aerobic conditions, and accumulation of both ppGpp and pppGpp was blocked under these conditions by pretreatment of the culture with rifampin or tetracycline. Growth and incorporation of [3H]guanosine, [3H]tymidine, [14C]succinate, and L-[35S]methionine into macromolecules were inhibited immediately upon exposure to air. The accumulation of ppGpp and pppGpp in B. thetaiotaomicron upon exposure to air may represent a novel signal for synthesis of these compounds.
Simplified preparation of mycoplasmas, an acholeplasma, and a spiroplasma for scanning electron microscopy
Authors: Gallagher J E , Rhoades K R
Published: January 2020

A simple, effective procedure was developed for scanning electron microscopic examination of mycoplasmas and similar organisms. Cultivation of several mycoplasmal species, an acholeplasma, and a spiroplasma in broth media in Leighton tubes with cover slips resulted in attachment of the organisms to the cover slips. The attached cells were easily processed for either scanning electron microscopy or light microscopy. By eliminating the need for centrifugation, which was used in previously described techniques, physical stress on the cell is minimized. The effects of different preparative procedures on the morphology of Mycoplasma gallisepticum are described.
Identification of the ftsA gene product
Authors: Lutkenhaus J F , Donachie W D
Published: January 2020

A nonsense mutation was identified in the essential cell division gene ftsA of Escherichia coli. A gamma-transducing phage was isolated which complemented this mutation. This phage programmed the synthesis of four bacterial proteins in UV-irradiated cells. By substituting the nonsense mutation for the ftsA+ allele in this transducing phage and comparing the proteins programmed by it in UV-treated Su+ and Su- cells, the product of the ftsA gene was identified as a protein with a molecular weight of 50,000.
Enzymes of agmatine degradation and the control of their synthesis in Klebsiella aerogenes
Authors: Friedrich B , Magasanik B
Published: January 2020

The degradation of agmatine to succinate by Klebsiella aerogenes occurs in five steps. The enzyme catalyzing the first step, agmatinase, is induced by agmatine. The enzymes catalyzing the second and third steps, putrescine aminotransferase and 4-aminobutyraldehyde dehydrogenase, are induced by putrescine and also by their product, 4-aminobutyrate. The enzymes catalyzing the fourth and fifth steps, 4-aminobutyrate aminotransferase and succinate semialdehyde dehydrogenase, are induced by 4-aminobutyrate. This compound also serves as gratuitous inducer of the catabolic acetylornithine aminotransferase. The formation of the enzymes responsible for agmatine degradation is regulated not only by induction, but also by catabolite repression and activation by glutamine synthetase.
Properties of two cyclic nucleotide-deficient mutants of Neurospora crassa
Authors: Rosenberg G , Pall M L
Published: January 2020

Studies on the crisp-1 (cr-1), cyclic adenosine 3',5'-monophosphate (cAMP)-deficient mutants of Neurospora crassa were undertaken to characterize the response of these mutants to exogenous cyclic nucleotides and cyclic nucleotide analogs. A growth tube bioassay and a radioimmune assay for cyclic nucleotides yielded the following results. (i) 8-Bromo cAMP and N6-monobutyryl cAMP but not dibutyryl cAMP are efficient cAMP analogs in Neurospora, stimulating mycelial elongation of the cr-1 mutants. Exogenous cyclic guanosine 3'5'-monophosphate (cGMP) also stimulates such mycelial elongation. (ii) Both cAMP levels and cGMP levels found in cr-1 mycelia are lower than those in wild type. However, the levels of both cyclic nucleotides are normal in conidia of cr-1. The data on cr-1 mycelia and those reported earlier in Escherichia coli (M. Shibuya, Y. Takebe, and Y. Kaziro (Cell 12:528-528, 1977) show a previously unexpected relationship between cAMP and cGMP metabolism in microorganisms. The semicolonial morphology of another adenylate cyclase-deficient mutant of Neurospora, frost, was not corrected by exogenous cyclic nucleotides or by phosphodiesterase inhibitors indicating that the frost morphology is probably not caused by low endogenous cAMP levels. The low adenylate cyclase activity and the abnormal morphology of frost may be related separately to the linolenate deficiency reported in the mutant.
Release of lipoteichoic acid from Streptococcus sanguis: stimulation of release during penicillin treatment
Authors: Horne D , Tomasz A
Published: January 2020

The spontaneous and the penicillin-stimulated release of water-soluble, glycerol-labeled polymers was compared in Streptococcus sanguis. In contrast to the spontaneous release occurring in exponentially growing or stationary-phase bacteria, penicillin-treated cells released the bulk of these polymers, and they were not replenished by synthesis during antibiotic treatment. Furthermore, a major portion of the extracellular polymers was characterized as acylated lipoteichoic acid.
Properties of the Bacillus subtilis spore coat
Authors: Pandey N K , Aronson A I
Published: January 2020

About 70% of the protein in isolated Bacillus subtilis spore coats was solubilized by treatment with a combination of reducing and denaturing agents at alkaline pH. The residue, consisting primarily of protein, was insoluble in a variety of reagents. The soluble proteins were resolved into at least seven bands by sodium dodecyl sulfate gel electrophoresis. About one-half of the total was four proteins of 8,000 to 12,000 daltons. These were relatively tyrosine rich, and one was a glycoprotein. There was also a cluster of proteins of about 40,000 daltons and two or three in the 20,000- to 25,000-dalton range. The insoluble fraction had an amino acid composition and N-terminal pattern of amino acids very similar to those of the soluble coat proteins. A major difference was the presence of considerable dityrosine in performic acid-oxidized preparations of insoluble coats. Coat antigen including a 60,000-dalton protein not present in extracts of mature spores was detected in extracts of sporulating cells by immunoprecipitation. This large antigen turned over in a pulse-chase experiment. Antibodies to either the array of 8,000- to 12,000-dalton coat polypeptides or to the larger coat proteins reacted with this 60,000-dalton species, suggesting a common precursor for many of the mature coat polypeptides. Spore coats seem to be assembled by processing of proteins and by secondary modifications including perhaps dityrosine formation for cross-linking.
Growth yields and fermentation balance of Bacteroides fragilis cultured in glucose-enriched medium
Authors: Frantz J C , McCallum R E
Published: January 2020

Bacteroides fragilis is an obligate anaerobic bacterium classified with the gram-negative, non-sporeforming bacilli and is the Bacteroides species most frequently isolated from human infections. In the present study, experiments were designed to investigate growth characteristics of B. fragilis in a complex medium. In a minimal defined medium, which was employed for comparison purposes, B. fragilis grew with a generation time of 2 h. Growth of the organism in glucose-enriched medium used in the present study was superior. Maximum generation time was 60 min. Total and viable cells (colony-forming units) were 8.9 x 10(9) and 2.1 x 10(9), respectively, at maximum measurable growth. The molar growth yield (Ym) was 51.5. Growth yields were found to reach a maximum 2 to 3 h before maximum growth and to vary with respect to the phase of growth. Estimates of the fermentation products indicated that glucose was the sole energy substrate. Major products included acetic acid, propionic acid, lactic acid, and succinic acid. Other products included ethyl alcohol, pyruvic acid, and fumaric acid. No attempt was made to recover CO2 or formic acid. The OR balances from two experiments were 0.013 and -0.093 and the respective carbon recoveries were 6.268 and 6.241. The results of the present study show that B. fragilis is capable of rapid rates of growth in vitro by using glucose as the sole energy source.
Relation between the adenylylation state of glutamine synthetase and the expression of other genes involved in nitrogen metabolism
Authors: Goldberg R B , Hanau R
Published: January 2020

We have partially characterized the biochemical parameters of glutamine synthetase from Klebsiella pneumoniae and have shown that the differential affinity of adenylylated and unadenylylated glutamine synthetase for adenosine diphosphate provides a convenient means of determining the adenylylation state. Using this assay procedure, we examined the relationship between the adenylylation state and the expression of other genes involved in nitrogen assimilation. We observed no correlation between the adenylylation state and the expression of histidase, glutamine synthetase, glutamate synthase, glutamate dehydrogenase, and urease in aerobic cultures.
Deoxyribonucleic Acid-Mediated Transformation of
Spirillum lipoferum
Authors: Mishra Ajit K. , Roy Pampa , Bhattacharya Sabita
Published: January 2020

The methodology for deoxyribonucleic acid-mediated transformation of
Spirillum lipoferum
to resistance to various antimicrobial agents is reported.
Association of tellurium resistance and bacteriophage inhibition conferred by R plasmids
Authors: Taylor D E , Summers A O
Published: January 2020

Concomitant resistance to tellurium compounds (Ter) and inhibition of coli-phage development (Phi) are properties mediated by many H2 incompatibility group R plasmids which have been isolated from diverse bacterial and geographic sources. Ter plasmids from tellurium-resistant bacteria that were isolated from sewage and industrial wastes also mediated phage inhibition. Of these Ter plasmids, three from Citrobacter freundii belonged to the H incompatibility group, whereas three from Klebsiella pneumoniae did not.
Addition of basic amino acids prevents G-1 arrest of nitrogen-starved cultures of Saccharomyces cerevisiae
Authors: Cooper T G , Britton C , Brand L , Sumrada R
Published: January 2020

Arginase-minus mutants of Saccharomyces cerevisiae were arrested in growth and accumulated at the unbudded G-1 stage of the cell cycle when starved for nitrogen. If, however, arginine was added to the culture medium at the time of starvation, growth ceased but the cells did not collect at the unbudded G-1 stage. We suggest that arginine addition prevented the cells from collecting at the G-1 stage by starving them for histidine and lysine, thereby inhibiting synthesis of proteins needed to complete the cell cycle.
Thermoresistant revertants of an Escherichia coli strain carrying tif-1 and ruv mutations: non-suppressibility of ruv by sfi
Authors: Otsuji N , Iyehara-Ogawa H
Published: January 2020

Spontaneous thermoresistant revertants were isolated from Tif1 Ruv- and Tif1 Ruv+ strains of Escherichia coli K-12. They were divided into five groups; backmutants to tif+ and recA structural gene mutants accounted for at least two of these groups. Mutations with an unconditional RecA- phenyotype were detected at a higher frequency in the Tif1 Ruv- strains (65%) than in the Tif1 Ruv+ strains (25%). A third group consisted of revertants exhibiting a RecA- phenotype at low temperature. Revertants with normal recombination ability and UV resistance, but with a thermosensitive defect in propagating lambda bio11 phage, were also isolated (group 4). The alleles responsible for this property were cotransducible with the srl gene, suggesting that they are located at the recA locus. Other revertants, which might carry lex, LEXB, or zab mutations, were UV sensitive and were able to propagate lambda bio11 phage (group 5). The sfi mutation, which suppresses filamentation in the Tif1 and UV-sensitive Lon- strains, does not restore UV resistance of the Ruv- mutant.
Alteration of the fatty acid composition of Escherichia coli by growth in the presence of normal alcohols
Authors: Sullivan K H , Hegeman G D , Cordes E H
Published: January 2020

The addition of normal alcohols in the series n-butanol to n-octanol to cultures of Escherichia coli ML308 grown on defined or lipid-free medium (at 17, 27, and 37 degrees C) caused an alteration in the fatty acid composition of this organism: the ratio of saturated to unsaturated fatty acids increased. Changes in the relative quantities of individual fatty acid species elicited by increasing concentrations of these alcohols were as follows: (i) myristic acid remained constant: (ii) palmitic acid increased; and (iii) the combined amount of palmitoleic plus cis-methylene hexadecanoic acids changed in a way which was reflected inversely by changes in the amount of cis-vaccenic acid. Comparable changes were not observed when cells were grown in the presence of n-nonanol and n-decanol in the concentration range tested. The changes observed upon addition of normal alcohols (n-butanol to n-octanol) paralleled, in part, the alterations in fatty acid composition observed when growth temperature was increased.
Meiotic mapping of yeast ribosomal deoxyribonucleic acid on chromosome XII
Authors: Petes T D
Published: January 2020

We have used meiotic mapping techniques to locate the position of the repeating ribosomal DNA (rDNA) genes of the yeast Saccharomyces cerevisiae. We found that the rDNA genes are located on the right arm of chromosome XII, approximately 45 map units centromere distal to the gene gal2. Together with mapping data from previous studies, this result suggests that the tandem array of rDNA genes contains at least two junctions with the non-rDNA of the yeast chromosome. In addition, we observed segregation patterns of the rDNA genes consistent with meiotic recombination within the rDNA gene tandem array in 3 of the 59 tetrads examined.
Modifiable chromatophore proteins in photosynthetic bacteria
Authors: Hui K M , Hurlbert R E
Published: January 2020

The chromatophores of Chromatium vinosum, as well as six other photosynthetic bacteria, contained two or more proteins which were insoluble when heated in the presence of sodium dodecyl sulfate (SDS) and 2-mercaptoethanol (beta-ME). When the chromatophores were dissolved at room temperature in SDS-beta-ME, these proteins were present in the SDS-polyacrylamide gel electrophoresis profiles, but when the samples were dissolved at 100 degrees C, they were absent or considerably diminished. When one-dimensional gels of chromatophores solubilized at room temperature were soaked in the SDS-beta-ME solution and heated to 100 degrees C and the gels were run in a second dimension, the proteins became immobilized in the original first-dimension gel, where they could be detected by staining. The two major proteins so affected in C. vinosum had apparent molecular weights of 28,000 and 21,000. The chromatophores of several other photosynthetic bacteria also contained predominant proteins between 30,000 and 19,000 molecular weight, which became insoluble when heated in the presence of SDS and beta-ME. In at least two of the species examined, these appeared to be reaction center proteins. The conditions causing the proteins to become insoluble were complex and involved temperature, SDS concentration, and the presence of sulfhydryl reagents. The chromatophores of four of the Chromatiaceae species and two strains of one of the Rhodospirillaceae species examined had a protein-pigment complex that was visible in SDS-polyacrylamide gel profiles of samples dissolved at room temperature but was absent in samples dissolved at 100 degrees C.
Arrays of hemispheric surface projections on Chlamydia psittaci and Chlamydia trachomatis observed by scanning electron microscopy
Authors: Gregory W W , Gardner M , Byrne G I , Moulder J W
Published: January 2020

Scanning microscopy of two strains of Chlamydia psittaci and four strains of Chlamydia trachomatis representative of the wide diversity in origin and behavior of members of the genus revealed patches of regular arrays of hemispheric projections on the surfaces of elementary bodies of all six strains. These distinctive and perhaps unique surface structure are probably present in all populations of chlamydiae.
Detection of FP Plasmids in Hospital Isolates of
Pseudomonas aeruginosa
Authors: Dean Harley F. , Royle Pam , Morgan Anthony F.
Published: January 2020

A simple method of detection of FP plasmids with chromosome-mobilizing ability in
Pseudomonas aeruginosa
has been developed.
Distribution of cell lengths in cultures of a lexA mutant of Escherichia coli K-12
Authors: Howe W E , Mount D W
Published: January 2020

Distributions of cell lengths in lexA+ and lexA mutant cultures during normal growth and under thymidine starvation conditions are presented. During normal growth lexA mutant cells were slightly shorter, on the average, than were lexA+ cells. lexA mutant cells were also shorter in comparison with lexA+ cells after a period of thymidine starvation. These results are consistent with the hypothesis that the lexA gene is involved in the coordination of cell division with DNA repair.
Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase
Authors: Sanzey B
Published: January 2020

Nalidixic acid (Nal), a drug which affects deoxyribonucleic acid gyrase activity, inhibits the expression of catabolite-sensitive genes: the three maltose operons, the lactose and galactose operons, and the tryptophanase gene. A correlation between the degree of sensitivity to Nal and that to catabolite repression has been observed. The expression of the threonine and tryptophan operons, insensitive to catabolite repression, is insensitive to Nal. The expression of the lacZ gene under the control of the IQ promoter is activated by Nal. Strains carrying a mutation in the nalA locus are resistant to these effects. Novobiocin, which inhibits the negative supercoiling activity of deoxyribonucleic acid gyrase, affects expression of the operons similarly to Nal. The involvement of promoters in Nal and novobiocin action, as well as a possible role of in vivo negative supercoiling in the selectivity of gene expression, are discussed.
Uptake of circular deoxyribonucleic acid and mechanism of deoxyribonucleic acid transport in genetic transformation of Streptococcus pneumoniae
Authors: Lacks S
Published: January 2020

Deoxyribonucleic acid (DNA) from the covalently closed circular DNA molecules of Pseudomonas phage PM2 was found to enter normally transformable cells of Streptococcus pneumoniae as readily as linear bacterial DNA. In a mutant of S. pneumoniae that lacks a membrane nuclease and is defective in DNA entry, as many molecules of PM2 DNA as of linear DNA were bound on the outside of cells at equivalent DNA concentrations. Bound DNA suffered single-strand breaks, but circular DNA with preexisting breaks was bound no better than closed circles. In the presence of divalent cations, DNA bound to cells of a leaky nuclease mutant showed double-strand breaks. At least the majority of PM2 DNA that entered normal cells was single stranded. These results are consistent with a mechanism for DNA entry in which DNA is first nicked on binding, then a double-strand break is formed by cleavage of the complementary strand, and continued processive action of the membrane nuclease facilitates entry of the originally nicked strand. Although the bulk of circular donor DNA appeared to enter in this way, the results do not exclude entry of a small amount of donor DNA in an intact form.
Catabolism of L-tyrosine in Trichosporon cutaneum
Authors: Sparnins V L , Burbee D G , Dagley S
Published: January 2020

Protocatechuic acid was a catabolite in the degradation of L-tyrosine by Trichosporon cutaneum. Intact cells oxidized to completion various compounds proposed as intermediates in this conversion, but they did not readily oxidize catabolites of the homogentisate and homoprotocatechuate metabolic pathways, which are known to function in other organisms. Cell extracts converted tyrosine first to 4-hydroxycinnamic acid and then to 4-hydroxybenzaldehyde and 4-hydroxybenzoic acid. The proposed hydration product of 4-hydroxycinnamic acid, namely, beta-(4-hydroxyphenyl)-hydracrylic acid, was synthesized chemically, and its enzymatic degradation to 4-hydroxybenzaldehyde was shown to be dependent upon additions of adenosine triphosphate and coenzyme A. The hydroxylase that attacked 4-hydroxybenzoate showed a specific requirement for reduced nicotinamide adenine dinucleotide phosphate. Protocatechuate, the product of this reaction, was oxidized by cell extracts supplemented with reduced nicotinamide adenine dinucleotide or, less effectively, with reduced nicotinamide adenine dinucleotide phosphate, but these extracts contained no ring fission dioxygenase for protocatechuate. Evidence is presented that the principal hydroxylation product of protocatechuate was hydroxyquinol, the benzene nucleus of which was cleaved oxidatively to give maleylacetic acid.
Structure of polyadenylic acid in the ribonucleic acid of Saccharomyces cerevisiae
Authors: Phillips S L , Tse C , Serventi I , Hynes N
Published: January 2020

Investigations of the structure of polyadenylic acid [poly(A)] in yeast have shown that there are two classes of poly(A) distinguished by size and kinetics of synthesis. Each class is found directly on the 3' end of messenger RNA. One class contains poly(A) molecules ranging from 60 to less than 20 nucleotides long. The longest molecules in this poly(A) class are the first to become labeled when cells are exposed to [3H]adenine. Label then appears in progressively smaller molecules. The second class of poly(A) is about 20 nucleotides long. The length homogeneity of this class and the presence in nuclear DNA of many copies of a polythymidylate sequence which is the same length suggests that this poly(A) is synthesized by transcription from DNA.
Lysogenic pneumococci and their bacteriophages
Authors: Bernheimer H P
Published: January 2020

About half of pneumococci recovered from pediatric patients and one-third of isolates from adult patients yielded bacteriophages active against one or more of four noncapsulated indicator strains of pneumococcus. Strains of capsular types most frequently causing pediatric infections were associated with lysogeny. Classical restriction-modification phenomena have been demonstrated in vivo with some of the temperate phages, and correlation of restriction with the presence of one or the other of the two known pneumococcal restriction endonucleases has been established. The temperate phages differ serologically and in several other characteristics from virulent pneumococcal phages previously described. All pneumococcal phages so far studied can be classified into a minimum of three serological groups.
Inhibition of
Escherichia coli
Biodegradative Threonine Dehydratase by Pyruvate
Authors: Park Linda S. , Datta Prasanta
Published: January 2020

Pyruvate inhibits
Escherichia coli
K-12 biodegradative threonine dehydratase activity by a mechanism distinct from product inhibition by α-ketobutyrate and catabolite inactivation by intermediary metabolites.
Behavior of Penicillin-Binding Proteins in
Escherichia coli
upon Heat and Detergent Treatments and Partial Purification of Penicillin-Binding Proteins 1A and 1B
Authors: Nakagawa Jun-Ichi , Matsuzawa Hiroshi , Matsuhashi Michio
Published: January 2020

Penicillin-binding proteins differ greatly in heat sensitivity and sensitivity to detergents. The partial purification of penicillin-binding 1A and 1B proteins from
Escherichia coli
is described.
Use of chlC-lac fusions to determine regulation of gene chlC in Escherichia coli K-12
Authors: Fimmel A L , Haddock B A
Published: January 2020

Gene fusions between the lac structural genes and the chlC locus were isolated, and the regulation of lac gene expression was studied. The fused lac genes were induced by nitrate anaerobically and repressed by the presence of oxygen.
Mechanism of export of colicin E1 and colicin E3
Authors: Jakes K S , Model P
Published: January 2020

The mechanism of export of colicins E1 and E3 was examined. Neither colicin E1, colicin E3, Nor colicin E3 immunity protein appears to be synthesized as a precursor protein with an amino-terminal extension. Instead, the colicins, as well as the colicin E3 immunity protein, appear to leave the cells where they are made, long after their synthesis, by a nonspecific mechanism which results in increased permeability of the producing cells. Induction of ColE3-containing cells with mitomycin C leads to actual lysis of those cells, as some time after synthesis of the colicin E3 and its immunity protein has been completed. Induction of ColE1-containing cells results in increased permeability of the cells, but not in actual lysis, and most of the colicin E1 produced never leaves the producing cells. Intracellular proteins such as elongation factor G can be found outside of colicinogenic cells after mitomycin C induction, along with the colicin. Until substantial increases in permeability occur, most of the colicin remains cell associated, in the soluble cytosol, rather than in a membrane-associated form.
Modulation of cytochrome biosynthesis in yeast by antimetabolite action of levulinic acid
Authors: Malamud D R , Borralho L M , Panek A D , Mattoon J R
Published: January 2020

Levulinic acid, a competitive inhibitor of delta-aminolevulinic acid dehydratase, was used to inhibit cytochrome biosynthesis in growing yeast cells. In Saccharomyces cerevisiae the antimetabolite acts by inhibiting delta-aminolevulinic acid dehydratase in vivo, causing an accumulation of intracellular delta-aminolevulinic acid and simultaneous decreases in all classes of mitochondrial cytochromes. Changes in cellular cytochrome content with increasing levulinic acid concentration suggested the existence of different regulatory patterns in S. cerevisiae and Candida utilis. In C. utilis, cytochrome a.a3 formation is very resistant to the antimetabolite action of levulinic acid. In this aerobic yeast, cytochrome c+c1 is the most sensitive to levulinic acid, and cytochrome b exhibits intermediate sensitivity.
Regulation of the biosynthesis of aminoacyl-transfer ribonucleic acid synthetases and of transfer ribonucleic acid in Escherichia coli. VI. Mutants with increased levels of glutaminyl-transfer ribonucleic acid synthetase and of glutamine transfer ribonucleic acid
Authors: Cheung A , Morgan S , Low K B , Söll D
Published: January 2020

Spontaneous revertants of a temperature-sensitive Escherichia coli strain bearing a thermolabile glutaminyl-transfer ribonucleic acid (tRNA) synthetase have been selected for growth at 45 degrees C. Among 10 revertants still containing the thermolabile enzyme, 2 interesting strains were found. One strain has a fivefold elevated level of the thermolabile glutaminyl-tRNA synthetase; the genetic locus, glnR, responsible for this effect maps at min 24, far from glnS, the structural gene of the enzyme. In the other strain the levels of tRNA Gln and several other tRNAs are twice as high as in the parental strain; the locus responsible, glnU, maps at min 59.5 on the E. coli map.
Levels of major proteins of Escherichia coli during growth at different temperatures
Authors: Herendeen S L , VanBogelen R A , Neidhardt F C
Published: January 2020

The adaptation of Escherichia coli B/r to temperature was studied by measuring the levels of 133 proteins (comprising 70% of the cell's protein mass) during balanced growth in rich medium at seven temperatures from 13.5 to 46 degrees C. The growth rate of this strain in either rich or minimal medium varies as a simple function of temperature with an Arrhenius constant of approximately 13,500 cal (ca. 56,500 J) per mol from 23 to 37 degrees C, the so-called normal range; above and below this range the growth rate decreases sharply. Analysis of the detailed results indicates that (i) metabolic coordination within the normal (Arrhenius) range is largely achieved by modulation of enzyme activity rather than amount; (ii) the restricted growth that occurs outside this range is accompanied by marked changes in the levels of most of these proteins; (iii) a few proteins are thermometer-like in varying simply with temperature over the whole temperature range irrespective of the influence of temperature on cell growth; and (iv) the temperature response of half of the proteins can be predicted from current information on their metabolic role or from their variation in level in different media at 37 degrees C.
Growth of Nocardia rhodochrous on acetylene gas
Authors: Kanner D , Bartha R
Published: January 2020

Soil sediment enrichment cultures yielded a coryneform bacterium capable of growing in a mineral salts solution with acetylene gas as its only source of carbon and energy. Based on morphological and physiological traits as well as on cell wall analysis, the bacterium was characterized as a strain of Nocardia rhodochrous. Maximal growth rates (generation time 2.7 to 3.0 h) on acetylene were obtained at 5 to 20% acetylene, 25 to 40% oxygen, pH 7.0 and 26 to 28 degrees C. Yields (grams of dry cells produced per gram of acetylene consumed) ranged between 90 and 110%. N. rhodochrous exhibits a growth factor requirement for the pyrimidine moiety of thiamine. Acetylene utilization is not an obligate trait, and a wide range of alternate carbon sources is utilized. Ethylene is neither produced nor consumed. The only previous report on acetylene utilization appeared in 1932. The Mycobacterium lacticola strain described in that report strongly resembles N. rhodochrous.
Pyrimidine dimer excision in a Bacillus subtilis Uvr- mutant
Authors: Hadden C T
Published: January 2020

A technique which allows the measurement of small numbers of pyrimidine dimers in the deoxyribonucleic acid (DNA) of cells of Bacillus subtilis irradiated with ultraviolet light has been used to show that a strain mutant at the uvr-1 locus is able to excise pyrimidine dimers. Excision repair in this strain was slow, but incision may not be rate limiting because single-strand breaks in DNA accumulate under some conditions. Excision repair probably accounted for a liquid-holding recovery previously reported to occur in this strain. Recombinational exchange of pyrimidine dimers into newly replicated DNA was readily detected in uvr-1 cells, but this exchange did not account for more than a minor fraction of the dimers removed from parental DNA. Excision repair in the uvr-1 strain was inhibited by a drug which complexes DNA polymerase III with DNA gaps. This inhibition may be limited to a number of sites equal to the number of DNA polymerase III molecules, and it is inferred that large gaps are produced by excision of dimers. Because the uvr-1 mutation specifically interferes with excision of dimers at incision sites, it is concluded that the uvr-1 gene product may be an exonuclease which is essential for efficient dimer excision.
Ribulose 1,5-bisphosphate carboxylase/oxygenase from Pseudomonas oxalacticus
Authors: Lawlis V B , Gordon G L , McFadden B A
Published: January 2020

Ribulose 1,5-bisphosphate carboxylase/oxygenase was purified by a rapid, facile procedure from formate-grown Pseudomonas oxalaticus. The electrophoretically homogeneous enzyme had specific activities of 1.9 mumol of CO2 fixed per min per mg of protein and 0.15 mumol of O2 consumed per min per mg of protein. The amino acid composition was similar to that of other bacterial sources of the enzyme. The molecular weights determined by sedimentation equilibrium and by gel filtration were 421,000 and 450,000, respectively. Upon sodium dodecyl sulfate electrophoresis of enzyme purified under conditions which would limit proteolysis, two types of large (L) subunits and two types of small (S) subunits were observed with apparent molecular weights of 57,000, 55,000, 17,000 and 15,000. By densitometric scans at two different protein concentrations the stoichiometry of the total large to total small subunits was 1:1, implying an L6S6 structure. Electron micrographs of the enzyme revealed an unusual structure that was inconsistent with a cubical structure. The enzyme had an unusually high Km for ribulose 1,5-bisphosphate (220 microM) and was strongly inhibited by 6-phosphogluconate in the ribulose 1,5-bisphosphate carboxylase assay (Ki = 270 microM). One, 5, and 12 days after purification the enzyme was half-maximally activated at 0.13 microM, 0.23 mM, and 0.70 mM CO2, respectively, at saturating Mg2+. At saturating CO2, enzyme 1 day afer purification responded sigmoidally to Mg2+ and was half-maximally activated by 0.85 mM Mg2+ in the absence of 6-phosphogluconate (Hill coefficient, h = 2.0) and by 0.19 mM Mg2+ in the presence of mM 6-phosphogluconate (h = 1.7).
Introduction of host-controlled modification and restriction systems of Bacillus subtilis IAM1247 into Bacillus subtilis 168
Authors: Shibata T , Ikawa S , Komatsu Y , Ando T , Saito H
Published: January 2020

Bacillus subtilis IAM1247 had two modification and restriction systems (Bsu1247I and Bsu1247II), the former producing an isoschizomer of PstI endonuclease. A transformant clone was isolated which had Bsu 168, BsuR, and Bsu1247I systems coexisting within a genome.
Isolation and characterization of two tryptophan biosynthetic enzymes, indoleglycerol phosphate synthase and phosphoribosyl anthranilate isomerase, from Bacillus subtilis
Authors: Hoch S O
Published: January 2020

Two of the enzymes responsible for tryptophan biosynthesis in Bacillus subtilis have been extensively purified. These proteins are indole-3-glycerol phosphate synthase and N-(5'-phosphoribosyl) anthranilate isomerase. By comparison to the non-differentiating enteric bacteria in which these two enzymes are fused into a single polypeptide, the isolation of the indoleglycerol phosphate synthase and phosphoribosyl anthranilate isomerase from B. subtilis has demonstrated that the two proteins are separate species in this organism. The two enzymes were clearly separable by anion-exchange chromatography without any significant loss of activity. Molecular weights were determined for both enzymes by gel filtration and sodium dodecyl sulfate-slab gel electrophoresis, and indicated that the indoleglycerol phosphate synthase is the slightly larger of the two proteins. The minimum molecular weight for indoleglycerol phosphate synthase was 23,500, and that for phosphoribosyl anthranilate isomerase was 21,800. Both enzymes have been examined as to conditions necessary to achieve maximal activity of their individual functions and to maintain that activity.
Escherichia coli mutant strain with altered expression of the tryptophan operon: isolation and preliminary characterization
Authors: Pouwels P H , Scholten H J
Published: January 2020

From a strain of Escherichia coli with two copies of the tryptophan (trp) operon and one copy of the lactose (lac) operon, under control of one of the trp regulatory elements, we have isolated a mutant which does not grow in a medium containing 19 amino acids, unless tryptophan is added, and which cannot ferment lactose. The apparent pleiotropic nature of the mutation(s) is indicated by the very slow growth of mutant bacteria on minimal-medium agar supplemented with glucose and tryptophan. The amount of the trp enzymes (anthranilate synthetase and tryptophan synthetase) and trp messenger ribonucleic acid is reduced several-fold in the mutant compared to the isogenic wild-type strain, whereas the enzymes tryptophanyl-transfer ribonucleic acid synthetase and glucose 6-phosphate dehydrogenase remain the same. The incorporation of radioactive label into pulse-labeled but not into stable ribonucleic acid is significantly lower. Our results suggest that in the mutant organism the control of transcription of some operons, including the trp operon, is modified. An alternative explanation is that mutant bacteria contain a ribonuclease with increased activity for some messenger ribonucleic acid species.