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E Englesberg

Publications and source records attributed to E Englesberg.

51 records · Page 3Linked to original sources
PubMed1971

Control of expression of the L-arabinose operon in temperature-sensitive mutants of gene araC in Escherichia coli B-r.

Expression of the l-arabinose operon in Escherichia coli B/r is dependent on the temperature of growth of the araC mutants reported in this paper. Analysis of these temperature-sensitive regulatory mutants indicates that both repressor and activator activities are thermolabile. The simplest model to explain the manner in which the operon is controlled is one suggesting that the regulatory gene, araC, codes for a protein which upon synthesis acts as a repressor molecule and prevents operon function. When inducer is added, the repressor undergoes a conformational shift and becomes an activator which switches on enzyme synthesis, provided the repressor concentration is reduced to a sufficiently low level in the cell. These data lend strong support to the model that both activities are the result of the same gene product.

Alleles↗
PubMed1971

Hyperinducibility as a result of mutation in structural genes and self-catabolite repression in the ara operon.

Mutations in gene araB producing an l-arabinose-negative phenotype cause either an increase (hyperinducible), decrease (polar), or have no effect at all on the inducible rate of expression of the l-arabinose operon. Fourteen araB gene mutants exhibiting such effects were shown to be the result of: nonsense, frameshift, or missense mutations. All missense mutants were hyperinducible, exhibiting approximately a twofold increase in rate of l-arabinose isomerase production. All frameshift and most nonsense mutants exhibited polar effect. One nonsense mutant was hyperinducible. The cis-dominant polar effect of nonsense and frameshift mutants (as compared to induced wild type) were more pronounced in arabinose-utilizing merodiploids and in araBaraC(c) double mutants where inducible and constitutive enzyme levels are respectively determined. On the other hand, in arabinose-utilizing merodiploids, missense mutations no longer exhibited hyperinducibility but displayed a wild-type level of operon expression. Increases in the wild type-inducible rate of expression of the operon were found when growth rate was dependent on the concentration of l-arabinose. Cyclic 3',5'-adenosine monophosphate also stimulated expression of the operon with the wild type in a mineral l-arabinose medium. These observations are explained on the basis that the steady-state expression of the l-arabinose operon OIBAD is dependent on the concentration of (i) l-arabinose, the effector of this system, which stimulates the expression of the operon, and (ii) catabolite repressors, produced from l-arabinose, which dampen the expression of the operon. We have termed the latter phenomenon "self-catabolite" repression.

Arabinose↗
PubMed1969

The L-arabinose operon in Escherichia coli B-r: a genetic demonstration of two functional states of the product of a regulator gene.

The product of the regulator gene araC in the L-arabinose gene complex exists in two functional states: P1, the repressor, and P2, the activator, presumably in equilibrium with each other, and with P1 and P2 attached to their respective controlling sites, araO, the operator, and araI, the initiator. The controlling sites are linked in the following order with respect to genes araB and araC: BIOC. Two C gene deletions (Delta719 and Delta766) serve to define the newly described araO site, and to place it adjacent to the left end of the C gene. We have suggested that deletion 719 deletes, in addition to the C gene, part or all of the araO site. Deletion 766 leaves the araO site intact. Complementation analysis employing stable merodiploids indicates that the repressor-operator site function is epistatic over the activator-initiator site function. It is necessary both for activator (P2) to be present and for repressor (P1) to be absent at their respective controlling sites (araI and araO) for full expression of the L-arabinose operon.

Arabinose↗
PubMed1969

L-arabinose binding protein from Escherichia coli B-r.

A protein which is capable of binding l-arabinose-1-(14)C has been isolated from l-arabinose-induced cultures of Escherichia coli B/r. Analysis for this l-arabinose-binding protein (ABP) in a number of l-arabinose-negative mutants suggests that the ABP is not coded for by any of the known genetic units of the l-arabinose complex yet is under the control of the regulator gene araC. The ABP has been purified and found to bind l-arabinose, d-fucose, d-xylose, and l-ribulose with decreasing affinities. The K(m) for l-arabinose is 5.7 x 10(-6)m. The molecular weight, as determined by equilibrium centrifugation, was found to be 32,000. The protein was observed to have many features that liken it to other recently isolated binding proteins that have been implicated in the active transport of small molecules.

Arabinose↗
PubMed1969

Arabinose-leucine deletion mutants of Escherichia coli B-r.

The control of ara gene expression was studied in mutants of Escherichia coli B/r containing deletions which fused the l-arabinose gene complex with the leucine operon (the normal gene order being araDABIOC...leuDCBAO). Complementation experiments with stable merodiploids showed that expression of ara genes cis to araC-leu deletions was controlled by the trans-acting product of the araC gene. Expression of ara genes cis to araB-leu deletions was under leucine control. These studies confirm the existence of a region between genes araC and araB essential for normal activator controlled expression of the ara structural genes. One deletion was characterized as an araO-leu deletion. Its effect on ara gene expression was unique in that ara genes were susceptible to potential regulation by both l-arabinose and leucine. These experiments suggest that two different species of messenger ribonucleic acid (mRNA) may be produced for the ara-leu region as a result of this deletion. One, under l-arabinose-activator control, is initiated in the l-arabinose region; the other, under leucine control, is initiated in the leucine region. The latter indicates that araI can be transcribed. Whether araI is transcribed in the former instance (mRNA made under activator control) remains to be established.

Arabinose↗
PubMed1965

Positive control of enzyme synthesis by gene C in the L-arabinose system.

Englesberg, Ellis (University of Pittsburgh, Pittsburgh, Pa.), Joseph Irr, Joseph Power, and Nancy Lee. Positive control of enzyme synthesis by gene C in the l-arabinose system. J. Bacteriol 90:946-957. 1965.-The l-arabinose gene complex consists of genes D, A, B, and C, linked in that order between the markers thr and leu, and an unlinked gene E. Genes D, A, B, and E are the structural genes for three inducible enzymes and permease, respectively. Gene C, with two mutant alleles, C(-) and C(c), is the regulatory gene exhibiting positive and negative control. C(-) mutants are deficient and C(c) mutants are constitutive for all three enzymes and permease. Complementation analysis, employing sexual merozygotes (A(-)C(+) x A(+)C(-)), with six different C(-) mutants, demonstrates that C(-) is recessive to C(+) (positive control). A total of 61 C(c) mutants, isolated as clones resistant to d-fucose inhibition, are linked to the leu ara region of the chromosome, and the 22 C(c) mutants that were analyzed in detail mapped within the C gene among the C(-) mutant sites. C(c) mutants produce various but coordinate levels of the two enzymes measured, and permease. Complementation analysis (A(-)C(c) x A(+)C(-), A(-)C(c) x A(+)C(+)) shows that C(c) is dominant to C(-) (positive control) and recessive to C(+) (negative control). Deletion mutants that extend into the C gene are l-arabinose permease-negative, thus supporting the positive regulatory role of the C gene. The name "activator gene" is proposed for genes of the C type to accentuate their positive role in gene expression. A working model consistent with these results is presented.

Arabinose↗