bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship Between Messenger Ribonucleic Acid and Enzyme Levels Specified by the Leucine Operon of
Escherichia coli
K-12
Authors: Davis Michelle Gall , Calvo Joseph M.
Published: January 2020

The levels of leucine-forming enzymes in
Escherichia coli
K-12 varied over a several thousand-fold range, depending upon conditions of growth. The highest levels were achieved by growing auxotrophs in a chemostat under conditions of leucine limitation. Under such conditions, enzyme levels were increased 45- to 90-fold relative to cells grown in minimal medium containing leucine (the latter values arbitrarily called 1). Leucine operon-specific messenger ribonucleic acid levels were elevated to about the same extent as enzyme levels in cells grown in a chemostat. Growth in media of greater complexity resulted in progressively lower levels of leucine-forming enzymes, reaching a value of less than 0.02 for growth in a medium containing tryptone broth and yeast extract. The levels of leucine operon-specified enzymes and messenger ribonucleic acid were also measured in strains containing about 25 copies of plasmid pCV1(ColE1-
leu
) per chromosome. For such strains grown in minimal medium, enzyme levels were proportional to the number of plasmids per cell. Furthermore, they followed the same trends as those described above upon derepression in a chemostat or upon repression following growth in rich media. Leucine messenger ribonucleic acid, measured both by pulse-labeling and hybridization-competition experiments, was roughly proportional to enzyme levels over this entire range. For a plasmid-containing strain grown in a chemostat under conditions of leucine limitation (about 100 plasmids per chromosome), about 27% of pulse-labeled ribonucleic acid was coded for by genes in or adjacent to the leucine operon, and 10% of the total protein was β-isopropylmalate dehydrogenase.
Autogenous regulation of the synthesis of glutamine synthetase in Klebsiella aerogenes
Authors: Bender R A , Magasanik B
Published: January 2020

We isolated an F' episome of Escherichia coli carrying the glnA+ gene from K. aerogenes and an F' episome of E. coli carrying the glnA4 allele from K. aerogenes responsible for the constitutive synthesis of glutamine synthetase. Complementation tests with these episomes showed that the glnA4 mutation (leading to the constitutive synthesis of active glutamine synthetase) was in the gene identified by mutations glnA20, glnA51, and glnA5 as the structural gene for glutamine synthetase. By using these merodiploid strains we were able to show that the glnA51 mutation lead to the synthesis of a glutamine synthetase that lacked enzymatic activity but fully retained its regulatory properties. Finally, we discuss a model that explains the several phenotypes associated with mutations such as glnA4 located within the structural gene for glutamine synthetase leading to constitutive synthesis of active glutamine synthetase.
Identification of a temperature-sensitive asparaginyl-transfer ribonucleic acid synthetase mutant of Escherichia coli
Authors: Yamamoto M , Nomura M , Ohsawa H , Maruo B
Published: January 2020

A temperature-sensitive mutant of Escherichia coli K-12 isolated previously (H. Ohsawa and B. Maruo, J. Bacteriol. 127:1157-1166, 1976) was found to have an alteration in asparaginyl-transfer ribonucleic acid synthetase. This alteration can account for the temperature-sensitive phenotype of the mutant. No evidence was obtained to support the previous suggestion that ribosomal protein S1 is altered in this mutant. Combined with the previous genetic studies, we conclude that the newly defined genetic locus, asnS, for the asparaginyl-transfer ribonucleic acid synthetase maps near pyrD at 21 min on the E. coli chromosome.
Membrane phospholipid synthesis and phenotypic correlation of an Escherichia coli pss mutant
Authors: Ohta A , Shibuya I
Published: January 2020

A pair of putatively isogenic pss(Ts) and pss+ (phosphatidylserine synthetase structural gene) strains was constructed and analyzed, together with the revertants, for the physiological consequences of cessation of the optimal synthesis of phosphatidylethanolamine (PE). Their in vivo and in vitro abilities to synthetize PE and the growth rates at different temperatures were determined. The rate of PE synthesis by OS2101 pss(Ts) was inversely related to the culture temperature. OS2101 in a low-salt broth medium stopped division and formed filamentous cells with declining viability upon the elevation of culture temperature from 27 to 42 or 44 degrees C, whereas the syntheses of deoxyribonucleic acid, ribonucleic acid, and protein were not affected. Proper concentrations of cations such as Na+, K+, NH4+, and Mg2+ or of sucrose could remedy the division and growth of OS2101 at the restrictive temperature without restoring normal PE synthesis. A remedial effect other than osmotic protection of these effectors and an adaptive regulatory mechanism for PE formation are suggested.
Unstable S-Adenosylmethionine synthetase in an ethionine-resistant strain of Neurospora crassa
Authors: Jacobson E S , Chen G S , Metzenberg R L
Published: January 2020

A pleitropic ethionine-resistant mutant of Neurospora contains an S-adenosylmethionine synthetase that is labile to heat and dialysis but exhibits normal kinetics for methionine and ethionine.
Microtubular structures in a stable staphylococcal L-form
Authors: Eda T , Kanda Y , Mori C , Kimura S
Published: January 2020

Microtubular structures, which were demonstrated as straight, dense-walled cylinders attached to cell membranes, were found in a stable staphylococcal L-form grown in the absence of antibiotic.
Identification of adenosine 3',5'-monophosphate in Mycobacterium smegmatis
Authors: Lee C H
Published: January 2020

Cyclic adenosine 3',5'-monophosphate isolated from Mycobacterium smegmatis cells was identified by thin-layer chromatography, stepwise conversion to adenosine 5'-monophosphate and adenosine, ultraviolet absorption spectrum, phosphate analysis, and detection by two relatively specific radioisotopic methods.
Cell cycle analysis of F'lac replication in Escherichia coli B/r
Authors: Finkelstein M , Helmstetter C E
Published: January 2020

The timing and control of replication of an F'lac plasmid was investigated in two substrains of Escherichia coli B/r lac/F'lac growing at a variety of rates. The cellular content of covalently closed circular F'lac deoxyribonucleic acid and the cellular mass at the time of F'lac replication both increased as a function of growth rate. The timing of plasmid replication during the division cycle was determined by measuring the inducibility of beta-galactosidase in cells of different ages in exponentially growing cultures. At all growth rates, the rate of induced beta-galactosidase synthesis increased in a step-wise fashion during the division cycle, indicating that the F'lac plasmid replicated at a discrete time in the cycle. At growth rates greater than one doubling per h, the cell age at F'lac replication was indistinguishable from the cell age at chromosomal lac+ replication in an isogenic F- parent. The ratio of plasmids to chromosomal origins decreased from about 0.7 to 0.4 between growth rates of 1.0 to 2.5 doublings per h. These observations are all consistent with replication of F'lac at about the same time in the division cycle as replication of the homologous chromosomal region at these growth rates. This similarity in timing of replication of homologous deoxyribonucleic acid regions was not evident in slower-growing cells.
Electron microscopy of yeastlike cell development from the microconidium of Histoplasma capsulatum
Authors: Garrison R G , Boyd K S
Published: January 2020

Fine details of the sequential morphological events occurring during transition of microconidia (spores less than 5 micrometer in diameter) to the yeastlike phase of Histoplasma capsulatum as seen in ultrathin section are described and illustrated by electron micrographs. Masses of microconidia were obtained when the fungas was grown on a garden soil extract medium. Spores were incubated under in vitro environmental conditions conducive for phase transition (an enriched medium at 37 degrees C). Within 48 h of incubation, the microconidia either germinated to give rise to a short mycelium or the germ tube process became a yeast mother cell without further extension. The wall of the yeast mother cell was thin and smooth, and its cytoplasmic content was ultrastructurally complex, consisting of numerous lipid bodies, vacuoles, glycogen-like deposits, and membrane systems. Within 96 h, the mother cell underwent multipolar budding to form simultaneously linear hyphal and/or ovate yeastlike daughter cells. During the transition, new cell wall materials of the germ tube, the mother cell, and yeastlike daughter cells arose by blastic action from the innermost layer(s) of the wall of the precursor form. Lomasome-like vesicles were often seen in association with areas of new cell wall formation. After organellar migration into and septation of the daughter cells, the yeast mother cell's cytoplasmic content underwent marked degenerative changes.
Cloning of Beneckea genes in Escherichia coli
Authors: Lamfrom H , Sarabhai A , Abelson J
Published: January 2020

Genes from Beneckea harveyi, a luminescent marine bacterium, were cloned in Escherichia coli. This was done by producing randomly sheared fragments of Beneckea DNA and inserting them into the EcoRI site of plasmid pMB9 by the adenine-thymine joining procedure. The hybrid plasmids were used to transform E. coli C600 SF8. Among the transformants selected for tetracycline resistance, one clone that appeared to complement a leucine tb mutation was identified. The transformants were screened for the presence of Beneckea 5S genes. Four of these clones were analyzed in detail by hybridization with 16S, 23S, and 4S Beneckea RNA. The observations suggest that the ribosomal genes in Beneckea are linked, but are present in a different order than those in E. coli.
Sequential changes in cell volume distribution during vitamin B12 starvation of Euglena gracilis
Authors: Shehata T E , Kempner E S
Published: January 2020

During vitamin B12 starvation of Euglena, a new peak appears in the cell volume distribution. Some cells are inhibited at a unique point in the cell cycle between the initiation of DNA synthesis and nuclear division. The mechanism of inhibition of other cells differs.
Association and dissociation of Neurospora crassa cytoplasmic ribosomes
Authors: Judewicz N D , Algranati I D
Published: January 2020

Dissociation and association factors of ribosomal particles were detected in extracts from Neurospora crassa at different stages of growth. The dissociation factor was easily released into the S100 supernatant fraction, whereas the association factor remained bound to the ribosomes.
Elevated cyclic AMP concentration in streptomycin-dependent Escherichia coli
Authors: Polglase W J , Iwacha D , Thomson M
Published: January 2020

Streptomycin-dependent Escherichic coli B and K-12 cultures, which have relaxed catabolite repression when grown to glucose-salts medium, have an elevated concentration of cyclic AMP.
Chemotaxis in Pseudomonas aeruginosa
Authors: Moench T T , Konetzka W A
Published: January 2020

A chemotaxis system for Pseudomonas aeruginosa was defined by using the method of Adler. Cells were attracted to compounds in the order ammonium chloride greater than amino acids greater than organic acids. Two sugars were assayed and elicited no response. Comparisons with other model systems are discussed.
Physiology of lysine permeases in Saccharomycopsis lipolytica
Authors: Beckerich J M , Heslot H
Published: January 2020

Two active lysine transport systems were detected in Saccharomycopsis lipolytica. No excretion of lysine out of the cells could be obtained, even by chasing with L-lysine or by poisoning with sodium azide. The kinetic properties of one of the permeases, the high-affinity lysine permease, were studied in detail. Its Km was 1.91 +/- 0.23 X 10(-5) M. It proved highly specific, the only potent competitive inhibitors being (i) arginine and its analogs L-canavanine and L-ornithine, and (ii) the lysine analogs L-5 aminoethylcysteine and L-4,5-transdehydrolysine. It is suggested that the high-affinity lysine permease is common to L-lysine, L-ornithine, and L-arginine. The other amino acids tested behaved as noncompetitive inhibitors. The variation of uptake during a growth cycle was studied on ammonia-rich, ammonia-poor, and ammonia-free media. In each case, the uptake exhibited a peak in the early exponential growth phase. No new permease activity was detected during the lag phase or the stationary phase. Ammonia ions competitively inhibited the uptake and also decreased the Vmax value.
Transfer of nitrogen fixation genes from a bacterium with the characteristics of both Rhizobium and Agrobacterium
Authors: Skotnicki M L , Rolfe B G
Published: January 2020

Strain T1K, reported to be Rhizobium trifolii strain T1 carrying the drug resistance plasmid RU-1drd, was able to transfer a cluster of nif+ genes to Escherichia coli K-12. Additional genetic material, resembling the gal-chlA region of E. coli, was also transferred from strain T1K. The segregation pattern of these transferred genes suggested that they were on a plasmid. Although strain TIK was able to nodulate red and white clover, it also formed very slow-growing galls on tomato stems and shared many physiological properties with Agrobacterium tumefaciens, to which it seemed more closely related than to R. trifolii. The R. trifolii hybrid T1 (R1-19drd), constructed by conjugation, did not share any of these properties of both A. tumefaciens. Thus, strain T1K appears to be a bacterium with properties of both A. tumefaciens and R. trifolii and with the capacity to transfer nif+ genes and other functions which it may have "cloned" from another bacterium such as Klebsiella.
Stability of ribosomes of Staphylococcus aureus S6 sublethally heated in different buffers
Authors: Hurst A , Hughes A
Published: January 2020

Cells of Staphylococcus aureus heated at 52 degrees C in magnesium-chelating buffers [pH 7.2, 50 mM potassium phosphate or 50 mM tris(hydroxymethyl)-aminomethane containing 1 mM ethylenediaminetetraacetic acid] leaked 260-nm absorbing material, shown to be RNA, and suffered destruction of their ribosomes. These cells did not regain their salt tolerance when repair was carried out in the presence of actinomycin D (5 microgram/ml). Cells similarly heated in magnesium-conserving buffers [pH 7.2, 50 mM tris(hydroxymethyl)aminomethane containing 10 mM MgCl2 or piperazine buffer] did not leak RNA, suffered no ribosomal damage when heated for 15 min, and recovered, at least partially, in the presence of actinomycin D. Ribosomal damage, is therefore, a consequence of Mg2+ loss and is not an effect of heat per se. Cells suspended in either Mg2+-chelating or Mg2+-conserving buffers lost salt tolerance to about the same extent during heating at 52 degrees C. Therefore, sublethal heat injury can not be attributed to ribosomal damage.
Transduction of plasmid determinants in Staphylococcus aureus and Escherichia coli
Authors: Ubelaker M H , Rosenblum E D
Published: January 2020

Buoyant density analysis of transducing lysates derived from Staphylococcus aureus and Escherichia coli indicated that phage particles bearing plasmid determinants contain a quantity of DNA equivalent to that found in the lytic particles. Transducing particles that bear plasmid determinants smaller than viral DNA must therefore contain a quantity of DNA in excess of a single plasmid genome. In the E. coli P1vir system, a dependence upon host-mediated recombination for the transduction of small plasmids, but not for large R factors or chromosomal genes, was observed. However, no evidence for the involvement of such functions in the transduction of S. aureus plasmids was obtained. Although the origin of the additional DNA in plasmid transducing particles has not been identified, circumstantial evidence has been presented in the staphylococcal system indicating that transducing particles carrying a small tetracycline plasmid are not formed by the wrapping of multiple copies of this plasmid DNA.
Mannitol and Fructose Catabolic Pathways of
Pseudomonas aeruginosa
Carbohydrate-Negative Mutants and Pleiotropic Effects of Certain Enzyme Deficiencies
Authors: Phibbs P. V. , McCowen S. M. , Feary T. W. , Blevins W. T.
Published: January 2020

Mutant strains of
Pseudomonas aeruginosa
PAO were isolated on the basis of their inability to utilize mannitol as sole carbon source for growth. Four linkage groups (I through IV) among these mutant strains were resolved by two-factor crosses using the general transducing phage F116, and the strains appeared to contain point mutations as evidenced by ability to give rise to spontaneous revertants with wild phenotype on mannitol minimal agar. Group I strains were affected only in ability to grow on mannitol; all were deficient in inducible mannitol dehydrogenase activity, and all but one were deficient in inducible mannitol transport activity. Fructokinase was induced in group I strains and in wild-type bacteria during growth in the presence of mannitol but not fructose, indicating the presence of a pathway specific for endogenously generated fructose. Cells grown on fructose contained phosphoenolpyruvate:fructose-1-phosphotransferase activity, and mannitol-grown cells contained a lower level of this activity. Group II mutants were deficient in constitutive phosphoglucoisomerase, failed to grow on mannitol, grew very slowly on glycerol and fructose, but grew normally on glucose and gluconate. Group III strains were deficient in both nicotinamide adenine dinucleotide- and nicotinamide adenine dinucleotide phosphate-linked glucose-6-phosphate dehydrogenase activities that reside in a single enzyme species. 6-Phosphogluconate appeared to be the inductive effector for this enzyme, which was not required for aerobic growth on glucose or gluconate. A single mannitol-negative mutant in group IV also failed to grow on glycerol and glucose, but no biochemical lesion was identified.
Nutrition of Myxococcus xanthus, a fruiting myxobacterium
Authors: Bretscher A P , Kaiser D
Published: January 2020

The minimal requirements for vegetative growth of Myxococcus xanthus have been sought. Isoleucine, leucine, and valine were required, and vitamin B12 was needed for the synthesis of methionine. Pyruvate was an excellent energy source and an efficient source of cellular carbon. Acetate, aspartate, glutamate, and most tricarboxylic acid cycle intermediates could also be utilized, but were less efficient sources of carbon and energy than was pyruvate. Many mono- and disaccharides were tested, but, in agreement with earlier results, none served as carbon-energy sources. A minimal medium (A1) has been devised that includes the essential amino acids and vitamin B12, with pyruvate and aspartate as carbon-energy sources. In this medium, M. xanthus could propagate indefinitely, and on it vegetative cells formed colonies with greater than 75% efficiency; hence, it is likely that no organic cofactors other than those present in A1 are required in more than trace amounts.
Mapping and characterization of the nad genes in Salmonella typhimurium LT-2
Authors: Foster J W , Moat A G
Published: January 2020

An ampicillin enrichment technique was used to isolate 39 nicotinic acid-requiring mutants of Salmonella typhimurium LT-2. Using interrupted-mating and transductional mapping procedures, three loci, designated nadA, nadB, and nadC, were identified. These loci mapped at 33, 82, and 6 min, respectively, on the S. typhimurium linkage map. The arrangement of the loci on the Salmonella linkage map corresponded closely to the nadA, nadB, and nadC loci on the Escherichia coli K-12 linkage map, indicating that the de novo pathway to nicotinamide adenine dinucleotide and the genes governing the enzymes involved in this pathway in S. typhimurium are very similar to those in E. coli. Evidence is also presented which indicates that the product of the nadC locus in S. typhimurium LT-2 is the enzyme quinolinic acid phosphoribosyltransferase. All nadC mutants of S. typhimurium secreted between 2 and 8 mumol of quinolinic acid per 100 ml of secretion medium. In addition, none of the nadC mutants isolated were able to grow in 10(-3) M quinolinic acid, whereas all nadA and nadB mutants of S. typhimurium grew well in the presence of quinolinic acid. Transductional crosses between nadB mutants provided evidence suggestive of more than one locus in the nadB region.
Fate of homospecific transforming DNA bound to Streptococcus sanguis
Authors: Raina J L , Ravin A W
Published: January 2020

The fate of [3H]DNA from Streptococcus sanguis str-r43 fus-s donors in [14C]S. sanguis str-s fus-r1 recipients was studied by examining the lysates prepared from such recipients at various times after 1 min of exposure to DNA. The lysates were analyzed in CsCl and 10 to 30% sucrose gradients; fractions from the gradients were tested for biological activity and sensitivity to nucleases, subjected to various treatments and retested for nuclease sensitivity, and run on 5 to 20% neutral and alkaline sucrose gradients. The results demonstrate that donor DNA bound to S. sanguis cells in a form resistant to exogenous deoxyribonuclease is initially single stranded and complexed to recipient material. Donor DNA can be removed from the complex upon treatment of the complex with Pronase, phenol, or isoamyl alcohol-chloroform. Within the complex, donor DNA is relatively insensitive to S1 endonuclease but can regain its sensitivity by treatment with phenol. With time the complex moves as a whole to associate physically with the recipient chromosome. After a noncovalent stage of synapsis, donor material is covalently bonded to and acquires the nuclease sensitivity of recipient DNA, while donor markers regain transforming activity and become linked to resident markers.
Potassium transport system of Rhodopseudomonas capsulata
Authors: Jasper P
Published: January 2020

Rhodopseudomonas capsulata required potassium (or rubidium or cesium as analogs of potassium) for growth. These cations were actively accumulated by the cells by a process following Michaelis-Menten saturation kinetics. The monovalent cation transport system had Km's of 0.2 mM K+, 0.5 mM Rb+, and 2.6 mM Cs+. The rates of uptake of substrates by the potassium transport system varied with the age of the culture, although the affinity constant for the substrates remained constant. The maximal velocity of uptake of K+ was lower in aerobically grown cells than in photosynthetically grown cells, although the Km's for K+ and for Rb+ were about the same.
Nutritional studies with Pseudomonas aeruginosa grown on inorganic sulfur sources
Authors: Schook L B , Berk R S
Published: January 2020

Pseudomonas aeruginosa was grown on a succinate-basal salts medium supplemented with various inorganic sulfur compounds as its sole source of sulfur. The organism was able to grow on the sodium salts of sulfide, thiosulfate, tetrathionate, dithionite, metabisulfite, sulfite, or sulfate, but not on those of dithionate. Analyses of the culture media after 24 h of growth indicated accumulation of sulfate from each inorganic sulfur source except sulfate. Manometric studies with resting cells obtained by growth on each of these sulfur sources yielded net oxygen uptake for all substrates except sulfite and dithionate. Similar results were obtained with extracts from these cells by spectrophotometric techniques. Thiosulfate oxidase activity appeared to be induced by growth on sulfide, thiosulfate, or tetrathionate, with little or no activity observed when cells were grown on inorganic sulfur sources of higher oxidative states. Metabisulfite oxidase appeared to be associated with growth on all inorganic sulfur compounds. Rhodanese activity appeared to be constitutively present, and its activity, observed only in soluble fraction, seemed independent of the growth medium employed. Thiosulfate and tetrathionate oxidase activities were studied in greater detail than some of the other sulfur oxidases, and both were found to be distributed between particulate and soluble fractions.
Assignment of tra cistrons to EcoRI fragments of F sex factor DNA
Authors: Achtman M , Skurray R A , Thompson R , Helmuth R , Hall S , Beutin L , Clark A J
Published: January 2020

We describe here the cloning of single EcoRI fragments from the tra region of F DNA using ColE1::Tn3 as vector. These plasmids, as well as the series of Skurray et al. (Proc. Natl. Acad. Sci. U.S.A. 73:64-68, 1976), have been used to refine the map positions of tra cistrons on the F factor as well as to define a new DNA transfer cistron, traM. The current map of the tra cistrons is presented. None of the known tra cistrons, with the exception of traG, straddles an EcoRI site. The EcoRI site at 82 kilobases splits the traG cistron into two portions, an operator-proximal portion necessary for F pilus synthesis and an operator distal portion involved in conjugation itself. The operon structure of the tra cistrons was reevaluated, and we found that traI is at least partially independent of transcription of the traA to traD operon.
Conjugative plasmids in Neisseria gonorrhoeae
Authors: Sox T E , Mohammed W , Blackman E , Biswas G , Sparling P F
Published: January 2020

A conjugation system initially discovered in beta-lactamase-producing gonococci mobilized small non-selftransmissible R plasmids encoding beta-lactamase (penicillinase) production into other gonococci, Neisseria, and Escherichia coli. This conjugation system was mediated by a separate selftransmissible plasmid of 23.9 X 10(6) daltons, pFA2. Conjugative plasmids capable of mobilizing R plasmids were also found in nearly 8% of the non-penicillinase-producing gonococci. These were similar to pFA2 in size, buoyant density, and restriction endonuclease digest patterns but were less efficient than pFA2 in mobilization of the penicillinase plasmid pFA3. The presence of conjugative plasmids in gonococci isolated before the appearance of penicillinase-producing strains indicates that a conjugation system for plasmid transfer predated the appearance of R plasmids in gonococci.
Murein biosynthesis during a synchromous cell cycle of Escherichia coli B
Authors: Mirelman D , Yashouv-Gan Y , Nuchamovitz Y , Rozenhak S , Ron E Z
Published: January 2020

The last stages of murein biosynthesis were studied in relation to the division cycle of Escherichia coli in cells synchronized by amino acid starvation (Ron et al., J. Bacteriol. 123:374--376, 1975). Murein synthesis and the activities of the D-alanine carboxypeptidase and transpeptidase were found to vary significantly during the cell cycle. Maximal synthesis and transpeptidation were observed immediately after cell division, whereas maximal D-alanine carboxypeptidase activity was detected before cell division. These results are in agreement with our earlier findings that before cell division there is a stage of increased hydrolysis of the C-terminal D-alanine moiety of newly synthesized murein strands.
Aspartokinase of Streptococcus mutans: purification, properties, and regulation
Authors: McCarron R M , Chang Y F
Published: January 2020

Aspartokinase from Streptococcus mutans BHT was purified to homogeneity and characterized. The molecular weight of the native enzyme was estimated to be 242,000 by gel filtration. Cross-linking of aspartokinase with dimethyl suberimidate and polyacrylamide gel electrophoresis of the amidinated enzyme in the presence of sodium dodecyl sulfate showed the enzyme to be composed of six identical subunits with a molecular wieght of 40,000. The optimal pH range for enzyme activity was 6.5 to 8.5. The apparent Michaelis-Menten constants for aspartate and ATP were 5.5 and 2.2 mM, respectively. The enzyme was stable within the temperature range of 10 to 35 degrees C. Aspartokinase was not feedback inhibited by individual amino acids, but was concertedly inhibited by L-lysine and L-threonine (93.5% inhibition at 10 mM each). The inhibition was noncompetitive with respect to aspartate (Ki = 10 mM) and mixed with respect to ATP. L-Threonine methyl ester and L-threonine amide were able to substitute for L-threonine in feedback inhibition, but the requirement for L-lysine uas strict. The feedback inhibitor pair protected the enzyme against heat denaturation. Aspartokinase synthesis was repressed by L-threonine; this repression was enhanced by L-lysine, but was slightly attenuated by L-methionine.
Recombination between compatible plasmids containing homologous segments requires the Bacillus subtilis recE gene product
Authors: Keggins K M , Duvall E J , Lovett P S
Published: January 2020

Plasmid pSL103 was previously constructed by cloning a Trp fragment (approximately 2.3 X 10(6) daltons) from restriction endonuclease EcoRI-digested chromosome DNA of Bacillus pumilus using the neomycin-resistance plasmid pUB110 (approximately 2.8 X 10(6) daltons) as vector and B. subtilis as transformation recipient. In the present study the EcoRI Trp fragment from pSL103 was transferred in vitro to EcoRI fragments of the Bacillus plasmid pPL576 to determine the ability of the plasmid fragments to replicate in B. subtilis. Endonuclease EcoRI digestion of pPL576 (approximately 28 X 10(6) daltons) generated three fragments having molecular weights of about 13 X 13(6) (the A fragment), 9.5 X 10(6) (B fragment, and 6.5 X 10(6) (C fragment). Trp derivatives of pPL576 fragments capable of autonomous replication in B. subtilis contained the B fragment (e.g., pSL107) or both the B and C fragments (e.g., pSL108). Accordingly, the B fragment of pPL576 contains information essential for autonomous replication. pSL107 and pSL108 are compatible with pUB110. Constructed derivatives of the compatible plasmids pPL576 and pUB110, harboring genetically distinguishable EcoRI-generated Trp fragments cloned from the DNA of a B. pumilus strain, exhibited relatively high frequency recombination for a trpC marker when the plasmid pairs were present in a recombination-proficient strain of B. subtilis. No recombination was detected when the host carried the chromosome mutation recE4. Therefore, the recE4 mutation suppresses recombination between compatible plasmids that contain homologous segments.
Suppressibility of recA, recB, and recC mutations by nonsense suppressors
Authors: Templin A , Margossian L , Clark A J
Published: January 2020

Mutations in the recA, recB, and recC genes of Escherichia coli K-12 were surveyed to ascertain whether or not they are suppressed by nonsense suppressors. Several mutations which map in or near the recA gene, but have not been called recA mutations, were also surveyed. An amber recB mutation, recB156, and an amber recC mutation, recC155, were isolated. One recB mutation, recB95, and four recC mutations, recC22, recC38, recC82, and recC83, were found to be suppressed by a UGA suppressor. In addition to the previously isolated amber recA mutation recA99, two other recA mutations, recA52 and recA123, were found to be suppressed by amber suppressor supD32 but not by supE44.
Architecture of the Outer Membrane of
Escherichia coli
III. Protein-Lipopolysaccharide Complexes in Intramembraneous Particles
Authors: van Alphen Loek , Verkleij Arie , Leunissen-Bijvelt Jose , Lugtenberg Ben
Published: January 2020

In a previous paper (A. Verkleij, L. van Alphen, J. Bijvelt, and B. Lugtenberg, Biochim. Biophys. Acta
466:
269-282, 1977) we have hypothesized that particles on the outer fracture face of the outer membrane ([Formula: see text]), with corresponding pits on the inner fracture face of the outer membrane ([Formula: see text]), consist of lipopolysaccharide (LPS) aggregates stabilized by divalent cations and that they might contain protein and/or phospholipid. In the present paper the roles of LPS, cations, and proteins in these [Formula: see text] particles are described more extensively, using a strain that lacks the major outer membrane proteins, b, c, and d (b
−
c
−
d
−
), and has a reduction in the number of [Formula: see text] particles of 75%. To study the role of divalent cations in the formation of [Formula: see text] particles, these b
−
c
−
d
−
cells were grown or incubated with Ca
2+
, Mg
2+
, or putrescine. The presence of Ca
2+
resulted in the appearance of many [Formula: see text] particles and [Formula: see text] pits. Mg
2+
and putrescine were less effective than Ca
2+
. Introduction of these particles was not accompanied by alterations in the relative amounts of LPS and cell envelope proteins. Ca
2+
treatment of a heptoseless derivative of a b
−
c
−
d
−
strain did not result in morphological changes. Incubation of Ca
2+
-treated cells with ethylenediaminetetraacetate caused the disappearance of the introduced particles as well as the release of more than 60% of the cellular LPS. These results strongly support the hypothesis that LPS is involved in the formation of [Formula: see text] particles and [Formula: see text] pits. The roles of various outer membrane proteins in the formation of [Formula: see text] particles were studied by comparing the freeze-fracture morphology of b
−
c
−
d
−
cells with that of cells which contain one of the outer membrane proteins b, c, d, and e or the receptor protein for bacteriophage lambda. The results showed that the presence of any of these five proteins in a b
−
c
−
d
−
background resulted in a large increase in the number of [Formula: see text] particles and [Formula: see text] pits, indicating that these proteins are, independent of each other, involved in the formation of [Formula: see text] particles and [Formula: see text] pits. The simplest explanation for the results is that in wild-type cells each particle consists of LPS complexed with some molecules of a single protein species, stabilized by either divalent cations or polyamines. It is hypothesized that the outer membrane of the wild-type cell contains a heterogeneous population of particles, of which 75% consists of protein b-LPS, protein c-LPS, and protein d-LPS particles. A function of these particles as aqueous pores is proposed.
Does cycloheximide-induced loss of phosphate uptake activity in Neurospora crassa reflect rapid turnover?
Authors: Beever R E , Burns D J
Published: January 2020

Addition of cycloheximide to Neurospora crassa germlings growing in liquid medium caused an exponential loss of phosphate uptake activity (half-life, ca. 2 h). No loss of activity resulted when germlings were resuspended, at the time of cycloheximide addition, in medium of a substantially lower phosphate concentration. It is concluded that the phosphate uptake systems are not subject to rapid turnover.
Isolation and characterization of motile Escherichia coli mutants resistant to bacteriophage chi
Authors: Icho T , Iino T
Published: January 2020

Four mutants of Escherichia coli that are resistant to the flagellotropic phage chi, but are motile, were isolated. When they were observed in liquid culture bylight microscopy, one mutant exhibited circular movement and another tumbled at high frequency on the surface of a glass slide. The remaining two mutants moved normally. None of these mutants adsorbed the wild-type strain of chi. P1 transduction revealed that the mutation sites of these four mutants were more than 97% contransducible with a site in hag, the structural gene for flagellin. When flagellins of these mutants were chromatographed on a diethylaminoethyl-cellulose column, two eluted slower and one eluted slightly faster than the flagellin of the parental strain. The other flagellin eluted at the same position as that of the parent. Host range mutants of phage chi, which could infect these bacterial mutants, were isolated.
Control of ammonium assimilation in Rhizobium 32H1
Authors: Ludwig R A
Published: January 2020

The symbiotic, nitrogen-fixing bacterium Rhizobium sp. 32H1 is a specialized ammonium producer during symbiosis. However, during free-living growth, Rhizobium 32H1 assimilates ammonium very poorly. Two pathways of ammonium assimilation exist in enteric bacteria. One is mediated by glutamate dehydrogenase, and the other is mediated by glutamine synthetase-glutamate synthase. The former pathway is altogether inoperative in Rhizobium 32H1; the latter pathway operates at a slow rate and is under strict negative control by ammonium itself. Rhizobium 32H1 glutamine synthetase activity is modulated by both repression-derepression and reversible adenylylation. For a biochemical process lacking an alternative pathway, such a regulatory pattern exacerbates the very process. This suggests that Rhizobium 32H1 restricts its own ammonium assimilation to maximize the contribution of fixed nitrogen to the host plant during symbiosis.
Position of the Extra Amino Acid Sequence in the Precursor Arabinose-Binding Protein ofEscherichia coli
Authors: Hardy S. J. S. , Randall L. L.
Published: January 2020

The extra amino acid sequence in the precursor arabinose-binding protein was shown to be either close to or at the N-terminus.
Kinetics of glucose repression of yeast cytochrome c
Authors: Zitomer R S , Nichols D L
Published: January 2020

The kinetics of glucose repression of cytochrome c synthesis was measured by a radioimmune assay. When 5 or 10% glucose was added to a derepressed culture, the rate of cytochrome c synthesis was reduced to the repressed level with a half-life of 2 min. The addition of 1 or 0.5% glucose repressed the rate of cytochrome c synthesis to the same level as high glucose concentrations but with a longer half-life of 3 min. Glucose repression had no effect on the stability or function of the cytochrome c protein. Cellular levels of active cytochrome c mRNA during glucose repression were measured by translation of total cellular polyadenylic acid-containing RNA and immunoprecipitation cytochrome c from the translation products. The results of these measurements indicate that glucose represses the rate of cytochrome c synthesis through a reduction in the level of translatable cytochrome c mRNA.
Molecular events during the release of delta-aminolevulinate dehydratase from catabolite repression
Authors: Mahler H R , Lin C C
Published: January 2020

Transfer of exponential-phase cells of Saccharomyces cerevisiae, previously grown in 2% glucose, to a derepression medium resulted in a prompt increase in the level of delta-aminolevulinate dehydratase, the rate-limiting enzyme of heme biosynthesis under these conditions. This derepression exhibited a lag of 35 min at 23 degrees C and required the participation of both RNA and protein syntheses. Dissection of the molecular events during this lag period disclosed that RNA synthesis, rnal gene function (messenger RNA transport from nucleus to cytosol), and initiation of protein synthesis were completed within less than 10, 18, and 24 min, respectively. The potential regulation of derepression by mitochondrial gene products and mitochondrial function was probed by means of a series of isogenic, respiration-deficient (rho-, pet-, and mit-) mutants; no such regulation was found.
Evidence that spo0A mutations are recessive in spo0A-/spo0A+ merodiploid strains of Bacillus subtilis
Authors: Trowsdale J , Chen S M , Hoch J A
Published: January 2020

The spo0A locus contains two types of closely linked mutations that block sporulation at stage 0: spo0A mutations (the most pleiotropic of the stage 0 markers) and spo0C mutations. It was previously thought that spo0A mutations were dominant in merodiploids of Bacillus subtilis, whereas spo0C mutations were recessive. We have shown that spo0A mutations were recessive when spo0A-/spo0A+ merodiploids were made in the genetic backgrounds of strains that were resistant to antibiotic produced by the wild-type strain. Reinvestigation of cultures of spo0A-/spo0A+ merodiploids constructed in the antibiotic-sensitive spo0A strain showed that they contained the spo0A allele at a low frequency, and they produced very few haploid Spo- segregants. These facts indicated that the cultures contained mostly homogenotic (spo0A+/spo0A+) cells. The reason for the poor survival of the spo0A-/spo0A+ merodiploids in the genetic background of the antibiotic-sensitive strain was not clear, but several possible explanations were given. It may have been related to the diploid state of other genes in the same merodiploid cells. The previous indication that spo0A mutations were dominant seems to have been based on the properties of a rare class of Spo- segregants that were probably selected in the presence of antibiotic.
Dimensions of Escherichia coli at various growth rates: model for envelope growth
Authors: Pierucci O
Published: January 2020

The duplication of Escherichia coli B/r is described based on two independent sequences, the replication of the genome and the growth of the envelope. It is proposed that (i) new envelope growth zones are activated coincident with the initiation of new rounds of chromosome replication; (ii) each zone is active in envelope synthesis from the time of its inauguration to the division which follows the completion of the round of chromosome replication (that is, for C + D min); and (iii) the rate of envelope synthesis at each site is constant, independent of the growth rate. Measurements of the surface areas of two E. coli B/r substrains growing at a variety of rates and during nutritional transitions are consistent with the predictions of the model.
Rate of major protein synthesis during the cell cycle of Caulobacter crescentus
Authors: Iba H , Fukuda A , Okada Y
Published: January 2020

The rate of major protein synthesis was examined during the synchronous differentiation of Caulobacter crescentus. Total cell proteins were pulse-labeled with [35S]methionine at different times in the swarmer cell cycle and analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. The rates of synthesis of total cell proteins and of about one-half of the individual major proteins examined increased through G1 and S periods but remained nearly constant during G2 period. The rates of synthesis of the other half of the individual major proteins either increased continuously throughout the swarmer cell cycle or doubled during S period. One stage-specific protein was also detected in late S period. For most of the major proteins examined, the rate of synthesis in the swarmer cell was less than that in the stalked cell. It seemed that, before the onset of G2 period, the Caulobacter cell was already able to synthesize each major protein at the additive rate of the two progeny cells. Compared to the stability of cellular proteins, the functional degradation rate of mRNA coding for individual major proteins was rapid, with half-lives of 0.4 to 5.8 min. It thus seems that the rate of major protein synthesis mainly reflects the transcriptional control of gene expression.
Autolysins and shape change in rodA mutants of Bacillus subtilis
Authors: Rogers H J , Taylor C
Published: January 2020

The biochemical phenotype of rodA mutants was not affected by the simultaneous presence in double mutants of the lyt gene which makes them 90 to 95% deficient in autolysin action. The only morphological effect of this deficiency on the expression of the rod gene was that both the rod and the coccal forms of the mutant failed to separate and grew as long chains of cells. Inhibition of protein synthesis stopped the increase in peptidoglycan that occurred when the growth temperature for the mutants was raised to 45 degrees C. These observations support the idea that a derepression of peptidoglycan synthesis occurs at this temperature. The increased amount of cellular peptidoglycan at the higher growth temperature is not likely to be the result of the concomitant switching off of autolytic enzyme action.
Identification of three genes controlling production of new outer membrane pore proteins in Escherichia coli K-12
Authors: Pugsley A P , Schnaitman C A
Published: January 2020

Escherichia coli K-12 strains carrying mutations in the ompB gene or double mutations in the tolF and par genes lack the major outer membrane proteins 1a and 1b. These strains are deficient in the transport of small hydrophylic compounds and are multiply colicin resistant. When revertants of these strains were sought, a number of extragenic pseudorevertants were obtained which produced new outer membrane proteins. These new proteins could be divided into three classes by differences in electrophoretic mobility on polyacrylamide gels, by differing specificities for transport of small molecules, and by the identification of three different genetic loci for genes controlling their production. These genetic loci are designated as nmpA (at approximately 82.5 min on the E. coli K-12 genetic map), nmpB (8.6 min), and nmpC (12 min). The new proteins produced in strains carrying nmpA, nmpB, or nmpC mutations did not cross-react with antiserum against a mixture of proteins 1a and 1b, or with antiserum against phage-directed protein 2. Production of the new membrane proteins restored sensitivity to some of the colicins.
Control of Escherichia coli growth by CO2
Authors: Repaske R , Clayton M A
Published: January 2020

Escherichia coli B dependence on CO2 for growth was demonstrated. At suboptimal CO2 concentrations the rate of growth was controlled by CO2 concentration.
Effects of antipain (a protease inhibitor) on respiration, viability, and excision of pyrimidine dimers in UV-irradiated Escherichia coli cells
Authors: Swenson P A , Schenley R L
Published: January 2020

The protease inhibitor antipain increases the effectiveness of UV irradiation on cessation of respiration and cell killing in Escherichia coli B/r cultures without affecting excision of pyrimidine dimers. The actions are similar to those caused by cyclic AMP in irradiated cultures.
ATP hydrolysis and synthesis by the membrane-bound ATP synthetase complex of Methanobacterium thermoautotrophicum
Authors: Doddema H J , Hutten T J , van der Drift C , Vogels G D
Published: January 2020

The membrane-bound ATP synthetase complex of Methanobacterium thermoautotrophicum showed maximum activity for ATP hydrolysis at pH 8, at temperatures between 65 and 70 degrees C, and at an ATP-Mg2+ ratio of 0.5. Anaerobic conditions were not prerequisite for enzyme activity. The enzyme showed a Km value for ATP of 2 mM, and activity was Mg2+ dependent; Mn2+, Co2+, Ca2+, and Zn2+ could replace Mg2+ to some extent. Other nucleoside triphosphates could be hydrolyzed. N,N'-dicyclohexylcarbodiimide inhibited ATP hydrolysis. A proton-motive force, artificially imposed by a pH shift or valinomycin, resulted in ATP synthesis in whole cells. The ATP synthetase complex of the thermophilic methanogenic bacterium is similar to those described in aerobic and anaerobic microorganisms.
Biosynthesis of uridine diphosphate N-acetylmannosaminuronic acid in rff mutants of Salmonella tryphimurium
Authors: Lew H C , Nikaido H , Mäkelä P H
Published: January 2020

In Salmonella typhimurium, three groups of genes located in rfb, rfe, and rff clusters are known to be involved in the biosynthesis of the enterobacterial common antigen. We found that enzymatic synthesis of uridine diphosphate N-acetylmannosaminouric acid, the activated form of a constituent sugar of the common antigen, followed the pathway previously described in Escherichia coli (N. Ichihara, N. Ishimoto, and E. Ito, FEBS Lett. 39:46--48, 1974). All of the six rff mutants tested, which fail to synthesize the common antigen, were deficient in one or both of the two enzymes needed for the synthesis of this sugar nucleotide from uridine diphosphate N-acetylglucosamine; these results established the physiological role of the pathway studied for the biosynthesis of N-acetylmannosaminuronic acid residues. The levels of these enzymes were not reduced in rfe mutants or rfb deletion mutants, although they produced no or only traces of the common antigen.