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C M Berg

Publications and source records attributed to C M Berg.

At least 55 records · Page 3Linked to original sources
PubMed1980

The ilvG gene is expressed in Escherichia coli K-12.

Three genes code for isozymes of acetohydroxy acid synthase (AHAS) in Escherichia coli K-12. To test the previously published supposition that one of them, ilvG, is silent in ilvO+ strains, we isolated mutants which had deletions of various lengths in the ilvGEDA operon. Some of these mutants have severely reduced levels of AHAS activity. We conclude that ilvG is expressed in ilvO+ strains but is deleted in these mutants. In addition, we find that AHAS II, the ilvG gene product, is sensitive to feedback inhibition by valine. We hypothesize that ilvO- mutations are ilvG frameshift mutations which render AHAS II valine resistant and enhance transcription of distal genes.

Acetolactate Synthase↗
PubMed1980

Salmonella typhimurium mutants defective in acetohydroxy acid synthases I and II.

An analysis of transposon-induced mutants shows that Salmonella typhimurium possesses two major isozymes of acetohydroxy acid synthase, the enzymes which mediate the first common step in isoleucine and valine biosynthesis. A third (minor) acetohydroxy acid synthase is present, but its significance in isoleucine and valine synthesis may be negligible. Mutants defective in acetohydroxy acid synthase II (ilvG::Tn10) require isoleucine, alpha-ketobutyrate, or threonine for growth, a mutant defective in acetohydroxy acid synthase I (ilvB::Tn5) is a prototroph, and a double mutant (ilvG::Tn10 ilvB::Tn5) requires isoleucine plus valine for growth.

Acetolactate Synthase↗
PubMed1980

Substrate channeling: alpha-ketobutyrate inhibition of acetohydroxy acid synthase in Salmonella typhimurium.

Excess alpha-ketobutyrate inhibited the growth of Salmonella typhimurium LT2 by inhibiting the acetohydroxy acid synthase-catalyzed synthesis of alpha-acetolactate (a valine precursor). As a result, cells were starved for valine, and both ilvB (encoding acetohydroxy acid synthase I) and ilvGEDA (ilvG encodes acetohydroxy acid synthase II) were derepressed. The addition of valine reversed the effects of alpha-ketobutyrate.

Acetolactate Synthase↗
PubMed1979

Escherichia coli K-12 auxotrophs induced by insertion of the transposable element Tn5.

The sites of insertion of the transposable kanamycin-neomycin resistance-determining element, Tn5, in the E. coli K-12 chromosome were assessed in a collection of over 300 auxotrophs. Although mutations in at least 45 different cistrons were obtained, the distribution of insertion sites was not completely random: proA or proB; cysG; and cysH, cysD or cysC mutants were found in excess.

Chromosome Mapping↗
PubMed1979

Physiological characterization of polar Tn5-induced isoleucine-valine auxotrophs in Escherichia coli K.12: evidence for an internal promoter in the ilvOGEDA operon.

The properties of 22 isoleucine-valine auxotrophs induced in Escherichia coli K-12 by the transposable element, Tn5, were characterized on the basis of growth requirements, cross-feeding behavior, and enzyme activity. Mutants defective in ilvA, ilvC, ilvD and ilvE were found. Mutation in ilvE were not completely polar on ilvD and ilvA enzyme activities (that is, ilvE mutants possessed a low constitutive level of expression of the enzymes coded by ilvD and ilvA), while mutations in ilvD were completely polar on ilvA enzyme activity. The data suggest that there is an internal promoter between the sites of Tn5 insertion in ilvE and ilvD.

DNA Transposable Elements↗
PubMed1977

Proline excretion in Escherichia coli: a comparison of an argD+ strain and a proline-excreting argD- derivative.

In an attempt to deduce the physiological basis of proline excretion in argD- strains of Escherichia coli K12, several properties of an argD+ (nonexcreting) and an argD- (excreting) derivative were compared. No difference was found in the transport or in the utilization of either proline or its immediate precursor, delta1-pyrroline-5-carboxylate (PCA). Furthermore, no differences were found in the physical or kinetic properties of partially purified preparations of the enzyme mediating the final step in proline biosynthesis, PCA reductase. The specific activity of PCA reductase was, however, consistently higher in crude extracts prepared from the argD- mutant.

Amidohydrolases↗
PubMed1977

Inhibition of amino acid transport in Escherichia coli by some beta-lactam antibiotics.

Among 10 antibiotics tested, cephaloridine and cephalothin showed the greatest inhibition of proline active transport. Ethylenediaminetetraacetate pretreatment of the cells did not enhance inhibition by any of these 10 antibiotics. The inhibition of active transport of 10 additional amino acids by cephaloridine and cephalothin was studied. Both antibiotics inhibited transport of three amino acids which, like proline, have transport systems resistant to osmotic shock; neither antibiotic inhibited transport of the remaining amino acids, including three with shock-resistant and four with shock-sensitive systems.

Amino Acids↗
PubMed1977

Partial purification and some properties of delta1-pyrroline-5-carboxylate reductase from Escherichia coli.

delta1-Pyrroline-5-carboxylate (PCA) reductase [L-proline:NAD(P)+5-oxidoreductase, EC 1.5.1.2] has been purified over 200-fold from Escherichia coli K-12. It has a molecular weight of approximately 320,000. PCA reductase mediates the pyridine nucleotide-linked reduction of PCA to proline but not the reverse reaction (even at high substrate concentrations). The partially purified preparation is free of competing pyridine nucleotide oxidase, PCA dehydrogenase, and proline oxidase activities. The Michaelis constant (Km) values for the substrate, PCA, with reduced nicotinamide adenine dinucleotide phosphate (NADPH) or NADH as cofactor are 0.15 and 0.14 mM, respectively. The Km values determined for NADPH and NADH are 0.03 and 0.23 mM, respectively. Although either NADPH or NADH can function as cofactor, the activity observed with NADPH is severalfold greater. PCA reductase is not repressed by growth in the presence of proline, but it is inhibited by the reaction end products, proline and NADP.

Chloromercuribenzoates↗
PubMed1976

Penicillin-induced lysis of Escherichia coli in medium that does not support sustained growth.

Exponentially growing cells of an Escherichia coli auxotroph that are washed and resuspended in buffer containing a carbon source undergo sufficient growth to render them susceptible to penicillin-induced lysis, even in the absence of the required metabolite. This lysis is probably responsible for the reduction in intracellular enzyme activity previously attributed to a direct effect of penicillin upon the enzyme.

Culture Media↗
PubMed1975

Effect of enrichment procedure upon auxotroph recovery in Escherichia coli K-12.

Proline-requiring auxotrophs are recovered preferentially after mutant enrichment procedures (e.g., penicillin) which perturb the cell envelope, but not after procedures (e.g., thymineless death) which affect other cellular targets. This probably stems from effects of penicillin and similar antibiotics upon proline metabolic and transport enzymes associated with the cell envelope.

Escherichia coli↗
PubMed1974

Proline excretion and indirect suppression in Escherichia coli and Salmonella typhimurium.

The last step in proline biosynthesis in Escherichia coli K-12, Salmonella typhimurium LT7, and a number of other enterobacterial isolates is regulated so that no proline is excreted, even if excess Delta(1)-pyrroline-5-carboxylate, the immediate precursor of proline, is added to a culture. In proline auxotrophs blocked at an early step in proline biosynthesis (proA or proB), reversion to prototrophy is often due to a mutation in the arginine pathway which diverts N-acetyl glutamate gamma-semialdehyde to proline synthesis, thus bypassing the proA or proB block. In such double mutants (proAB, argD), the last step in proline synthesis appears to be unregulated, since proline is excreted. Feedback inhibition and repression of the arginine pathway overcomes indirect suppression (restoring the Pro(-) phenotype), but proline regulation is not restored; double mutants still excrete proline when fed Delta(1)-pyrroline-5-carboxylate exogeneously. A new class of proline analogue-resistant mutant, due to mutation at argD, is also described.

Arginine↗
PubMed1973

Thymineless death in polA+ and polA- strains of Escherichia coli.

The extreme sensitivity of polA(-) cells to thymineless death is due, primarily, to the absence of an extended lag prior to the commencement of death. Once thymineless death has commenced, the rate in polA(-) cells is only slightly faster than in polA(+) cells.

Culture Media↗
PubMed1971

Differential recovery of auxotrophs after penicillin enrichment in Escherichia coli.

Various auxotrophs are recovered from a penicillin enrichment cycle with differing efficiencies. Reconstruction experiments indicate that, under starvation conditions in the presence of penicillin, most auxotrophs undergo some death, whereas prolineless mutants are virtually immune to penicillin-induced killing.

Bacteriological Techniques↗
PubMed1971

Auxotroph accumulation in deoxyribonucleic acid polymeraseless strains of Escherichia coli K-12.

Spontaneous auxotrophs are found with high frequency in several strains of Escherichia coli K-12 deficient in Kornberg deoxyribonucleic acid polymerase. These include amino acid-, vitamin-, purine-, and pyrimidine-requiring strains. Although this was suggestive evidence that these strains might be mutators, reconstruction experiments demonstrate that auxotrophs possess a selective advantage over prototrophs in the same culture. Thus, despite the high frequency of auxotrophs in polymerase-deficient strains, it is not yet clear whether they have elevated mutation rates.

Adenine↗