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Regulation of the feruloyl esterase (faeA) gene from Aspergillus niger.

Feruloyl esterases can remove aromatic residues (e.g., ferulic acid) from plant cell wall polysaccharides (xylan, pectin) and are essential for complete degradation of these polysaccharides. Expression of the feruloyl esterase-encoding gene (faeA) from Aspergillus niger depends on D-xylose (expression is mediated by XlnR, the xylanolytic transcriptional activator) and on a second system that responds to aromatic compounds with a defined ring structure, such as ferulic acid and vanillic acid. Several compounds were tested, and all of the inducing compounds contained a benzene ring which had a methoxy group at C-3 and a hydroxy group at C-4 but was not substituted at C-5. Various aliphatic groups occurred at C-1. faeA expression in the presence of xylose or ferulic acid was repressed by glucose. faeA expression in the presence of ferulic acid and xylose was greater than faeA expression in the presence of either compound alone. The various inducing systems allow A. niger to produce feruloyl esterase not only during growth on xylan but also during growth on other ferulic acid-containing cell wall polysaccharides, such as pectin.

Aspergillus niger↗

Regulatory properties of araC(c) mutants in the L-arabinose operon of escherichia coliB/r.

Merodiploids containing a high-constitutive and a low-constitutive araC(c) allele were assayed for constitutive expression of the ara operon. Low-constitutive araC(c) alleles either were unable to repress the constitutive rate of ara operon expression exhibited by by high-constitutive araC(c) alleles or achieved a partial repression of the high-constitutive rate of operon expression. Either mutation to a low-constitutive araC(c) mutant resulted in a partial or complete loss of repressor function, or subunit mixing between the two araC(c) mutant proteins resulted in a partial or complete dominance of the high-constitutive araC(c) allele. Five of the six araC(c) alleles tested allowed a partial induction of the ara operon in cya crp background. In general, a higher level of ara operon induction was achieved in the cya crp background by high araC(c) alleles than by low araC(c) alleles. Furthermore, several araC(c) mutants exhibited decreased sensitivity to catabolite repression, particularly in the presence of inducer. The results suggest a model in which certain araC(c) gene products can achieve ara operon induction in the presence of either arabinose (inducer) or catabolite activator protein-cyclic adenosine monophosphate, whereas the wild-type araC gene product requires the presence of both of these factors for operon expression.

Alleles↗

Catabolite-repression-like phenomenon in Rhizobium meliloti.

We report a phenomenon similar to catabolite repression in Rhizobium meliloti. Succinate, which allows the highest observed rate of growth of R. meliloti, caused an immediate reduction of beta-galactosidase activity when added to cells growing in lactose. A Lac- mutant was unaltered in nodulation and nitrogen fixation capacities, but a pleiotropic mutant deficient in several catabolic properties was unable to produce effective nitrogen-fixing nodules.

Cyclic AMP↗

Regulation of C4-dicarboxylic acid transport in Bacillus subtilis.

Various mutant strains of Bacillus subtilis were used to study the induction and regulation of the transport of tricarboxylic acid cycle C4-dicarboxylates. L-Malate was the only physiological inducer and bromosuccinate was a gratuitous inducer of dicarboxylic acid transport in a succinic dehydrogenase deficient mutant. Several mutants were isolated with alterations in the ability to transport dicarboxylic acids. One of these (WK6) was defective in the ability to take up succinate when grown on glutamate minimal medium, whereas another (WK1) was inducible to high levels by extremely low levels of L-malate. Alpha-Ketoglutarate dehydrogenase mutants were unable to take up dicarboxylates because of repression of transport by glutamate and (or) alpha-ketoglutarate. A mutation which resulted in increased levels of alpha-ketoglutarate dehydrogenase partially overcame this inhibition. Glutamate did not prevent the induction of dicarboxylic acid transport by L-malate in succinic dehydrogenase mutants but was markedly inhibitory in alpha-ketoglutarate dehydrogenase mutants.

Aspartic Acid↗

Reduced level of ATF is correlated with transcriptional repression of DNA topoisomerase II alpha gene during TPA-induced differentiation of HL-60 cells.

DNA topoisomerase II is a marker for the proliferation state of mammalian cells in culture, and the protein levels are markedly higher in exponentially growing cells than quiescent cells and can be downregulated by growth of the cells at high density and serum starvation. Correlation between ATF and TPA-repressed DNA topoisomerase II alpha (Topo II alpha) mRNA has been investigated during TPA-induced differentiation of HL-60 cells. Topo II alpha mRNA and unknotting activity were reduced at 24 hours in TPA-treated HL-60 cells. The level of Topo II alpha mRNA and the activity were gradually decreased in proportion to the concentration of TPA. Two DNA-protein complexes were formed by DNA mobility shift assay when ATF-binding site was incubated with nuclear extract prepared from TPA-free HL-60 cells, and the amount of ATF was vanished after TPA treatment. TPA-repressed Topo II alpha mRNA and ATF levels were partially restored after pretreatment of staurosporin. These results suggest that the reduced level of ATF may be important to the transcriptional repression of Topo II alpha gene during TPA-induced differentiation in HL-60 cells and related to protein kinase C signal pathway.

Activating Transcription Factors↗

[Acetic acid, a catabolite repressor of cholinesterase synthesis by an Arthrobacter simplex culture].

Acetic acid was found to repress cholinesterase synthesis in the cells of Arthrobacter simplex var. cholinesterasus even at very low concentrations (0.1%). The repression is very stable. It is not eliminated by glucose or an organic acid of the Krebs cycle being added to the medium with acetic acid. The combination of acetic and butyric acids decreases the repression but does not eliminate it. The kinetics of cholinesterase synthesis was different in the cells grown on the medium with acetic acid and the cells cultivated on the medium with acetic acid and glucose, then washed and transferred to a fresh growth medium with glucose and acetylcholine as the sources of carbon.

Acetates↗

SIP1 is a catabolite repression-specific negative regulator of GAL gene expression.

The yeast Snf1p kinase is required for normal expression of many genes involved in utilization of non-glucose carbon. Snf1p is known to associate with several proteins. One is Sip1p, a protein that becomes phosphorylated in the presence of Snf1p and thus is a candidate Snf1p kinase substrate. We have isolated the SIP1 gene as a multicopy suppressor of the gal83-associated defect in glucose repression of GAL gene expression. Multicopy SIP1 also suppressed the gal82-associated defect in glucose repression, suggesting that SIP1, GAL83 and GAL82 function interdependently. Multicopy SIP1 gene reduces GAL1, GAL2, GAL7 and GAL10 gene expression three- to fourfold in cells grown in the presence of glucose but has no effect in cells grown on nonrepressing carbon. Sip1-deletion cells exhibited a two- to threefold increase in GAL gene expression compared to wild-type cells when grown on glucose. These studies show that SIP1 is a catabolite repression-specific negative regulator of GAL gene expression. Northern analysis revealed two SIP1 transcripts whose relative abundance changed with carbon source. Western blots revealed that Sip1p abundance is not markedly affected by carbon source, suggesting that Sip1p may be regulated post-translationally.

AMP-Activated Protein Kinases↗

Enhancement of isoleucine hydroxamate-mediated growth inhibition and improvement of isoleucine-producing strains of Serratia marcescens.

Growth inhibition by isoleucine hydroxamate in Serratia marcescens was significantly enhanced by adding valine plus leucine and by using glycerol as the carbon source. Isoleucine hydroxamate-resistant mutants were isolated under conditions in which growth inhibition was enhanced. One of the mutants, strain GIHVLr2179, lacked both feedback inhibition and repression of threonine deaminase. An alpha-aminobutyric acid-resistant mutant derived from strain GIHVLr2179, strain GIHVLAr2795, produced 12 mg of isoleucine per ml in the medium containing glucose and urea as carbon and nitrogen sources (a twofold increase over prior reports). This strain had increased activities of threonine deaminase, acetohydroxy acid synthase, aspartokinase, and homoserine dehydrogenase.

Acetolactate Synthase↗

Phosphoinositides are involved in control of the glucose-dependent growth resumption that follows the transition phase in Streptomyces lividans.

The interruption of the sblA gene of Streptomyces lividans was previously shown to lead to relief of glucose repression of the normally strongly glucose-repressed alpha-amylase gene. In addition to this relief, an early entry into stationary phase was observed when cells were grown in a minimal medium containing glucose as the main carbon source. In this study, we established that this mutant does not resume growth after the transition phase when cultured in the complex glucose-rich liquid medium R2YE and sporulates much earlier than the wild-type strain when plated on solid R2YE. These phenotypic differences, which were abolished when glucose was omitted from the R2YE medium, correlated with a reduced glucose uptake ability of the sblA mutant strain. sblA was shown to encode a bifunctional enzyme possessing phospholipase C-like and phosphoinositide phosphatase activities. The cleavage of phosphoinositides by SblA seems necessary to trigger the glucose-dependent renewed growth that follows the transition phase. The transient expression of sblA that takes place just before the transition phase is consistent with a regulatory role for this gene during the late stages of growth. The tight temporal control of sblA expression was shown to depend on two operator sites. One, located just upstream of the -35 promoter region, likely constitutes a repressor binding site. The other, located 170 bp downstream of the GTG sblA translational start codon, may be involved in the regulation of the degradation of the sblA transcript. This study suggests that phosphoinositides constitute important regulatory molecules in Streptomyces, as they do in eukaryotes.

Bacterial Proteins↗

[Changes in proteinase A and B activity during glucose repression in the yeast Saccharomyces cerevisiae].

The divisible and indivisible yeast S. cerevisiae were subjected to glucose repression by increasing the glucose content in the growth medium up to 10%. Under these conditions the total activities of both proteinases did not change significantly, while those of free (i.e. not bound to natural inhibitors) proteinases were increased manyfold. This effect is probably due to liberation of the proteinases from the vacuoles and digestion of cytosolic proteinase inhibitors.

Aspartic Acid Endopeptidases↗

Inhibition of polyamine accumulation and deoxyribonucleic acid synthesis in regenerating rat liver.

Repeated injections of 1,3-diaminopropane into rats after partial hepatectomy caused a repression-type inhibiton of liver ornithine decarboxylase (EC 4.1.1.17) and totally prevented the marked increases in liver putrescine and spermidine concentrations that normally occur in response to partial hepatectomy. The inhibition of polyamine synthesis by diaminopropane was accompanied by a profound decrease (about 80%) in the synthesis of DNA in the regenerating rat liver without any changes in the synthesis of RNA and total liver protein.

Animals↗

[Fatty acid makeup of Escherichia coli cells with repressed and derepressed phosphohydrolase biosynthesis].

Fatty acid composition of the cells of Escherchia coli wild strains K-10, and K-12 and the mutants of the regulatory genes for alkaline phosphatase was studied in conditions of repression and derepression of biosynthesis of phosphohydrolases. Derepression of phosphohydrolases was not accompanied with specific changes in fatty acid composition of the cells. An increase in the content of cyclopropanic acid in conditions of phosphorus deficiency and a decrease in the level of unsaturated fatty acids are related to deceleration of growth of the cells in these conditions.

Enzyme Repression↗

Developmental repression of growth and gene expression in Aspergillus.

Asexual reproductive development can be initiated in Aspergillus nidulans in the presence of excess nutrients through artificial induction of the developmental regulatory genes brlA or abaA by fusing the genes to the promoter from the alcohol dehydrogenase I gene (alcA) and culturing cells in the presence of an inducing alcohol. Artificially induced development completely inhibits growth and represses expression of the endogenous alcA gene and the coordinately controlled aldehyde dehydrogenase gene (aldA). Repression of alcA and aldA expression probably occurs at both the transcriptional and posttranslational levels. We propose that developmental induction results in a generalized metabolic shutdown, leading to an inability of cells to acquire nutrients from the growth medium. Self-imposed nutrient limitation could reinforce the primary developmental stimulus and ensure progression through the asexual reproductive pathway.

Alcohol Dehydrogenase↗

Effects of organic matter on the growth of Thiobacillus intermedius.

London, Jack (University of California, Los Angeles), and Sydney C. Rittenberg. Effects of organic matter on the growth of Thiobacillus intermedius. J. Bacteriol. 91:1062-1069. 1966.-Yeast extract, glucose, glutamate, and other organic materials stimulate the rate and extent of growth of Thiobacillus intermedius in thiosulfate broth. Growth did not occur in glucose or glutamate mineral salts medium in the absence of thiosulfate, although a stable variant was obtained which grows on yeast extract alone. Cells harvested from media supplemented with organic matter have a reduced rate of thiosulfate oxidation (20 to 30% of autotrophic), oxidize the organic supplement, and have an additive rate of oxidation in the presence of both the organic substrate and thiosulfate. Carboxydismutase synthesis is repressed, and the incorporation of bicarbonate carbon into cell material is almost completely eliminated by the presence of organic matter in the growth medium. It is concluded that the availability of organic matter eliminates the autotrophic assimilatory mechanisms of T. intermedius but not its autotrophic energy-generating system. The data are discussed in relation to the existence of "obligate" chemoautotrophic bacteria.

Bicarbonates↗

[Effect of growth medium composition of glucoamylase and glycosyltransferase activity of Endomyces fibuliger].

Production of glucoamylase and glycosyltransferase by Endomyces fibuliger was found to depend on sources of carbon and nitrogen nutrition. Starch at a concentration above 0.5% in the medium stimulated biosynthesis of glycosyltransferase but inhibited production of glucoamylase by End. fibuliger 20-9. The rate of growth of the micro-organism increased by a factor of 3.3 with an increase of starch concentration from 0.5 to 6%. Synthesis of glycosyltransferase was repressed by glucose, lactose, sucrose and maltose. Synthesis of glucoamylase was repressed by lactose, sorbose and galactose. Synthesis of glycosyltransferase was stimulated by xylose, sorbose and galactose. Production of glucoamylase was stimulated by xylose and arabinose. Growth of the culture and synthesis of glucoamylase and maltase in the cultural broth were stimulated by an increase in the concentration of maize extract. Biosynthesis of glucoamylase and glycosyltransferase was stimulated by NH4H2PO4.

Ascomycota↗

Multiple copies of SUC4 regulatory regions may cause partial de-repression of invertase synthesis in Saccharomyces cerevisiae.

Transformation to generate multiple copies of regulatory DNA sequences has been used to study the interactions between regulatory proteins and their target sequences, since a high copy number of these sequences may titrate trans-acting regulatory proteins. We have analyzed the synthesis of invertase in yeast strains carrying different SUC genes transformed with the multiple-copy plasmid pSH143, a derivative of pJDB207 containing the promoter and upstream regulatory sequences of SUC4. The results obtained seem to be strain dependent. Under repressing conditions a high copy number of SUC4 promoter regions may cause increased expression of the invertase genes resulting in the synthesis of external glycosylated protein. A similar result was obtained under de-repressing conditions since transformants from some strains showed higher levels of activity. These results suggest that transcriptional regulatory (negative) factors may become limiting when the copy number of their target DNA sequences is increased. This effect may depend on the amount of active repressor molecules as well as on their affinity for SUC4 upstream sequences. This is discussed on the basis of the nucleotide sequences of SUC promoters.

Base Sequence↗

Genes involved in the uptake and catabolism of gluconate by Escherichia coli.

The isolation and properties of a mutant of Escherichia coli K12 that is totally unable to take up and utilize gluconate are described. Genetical analysis shows this phenotype to be associated with two lesions. One phenotype, designated GntM-, is the result of a mutation in a gene co-transducible with malA; the other, designated GNTS-, is the result of a mutation in a gene (GntS) co-transducible with fdp. The GntS--phenotype differs little from that of wild-type cells, but GntM- GntS+ organisms grow on gluconate only after a prolonged lag and form a gluconate uptake system that is strongly repressed by pyruvate. Moreover, such GntM- mutants readily give rise to further mutants that form a gluconate uptake system, gluconate kinase and 6-phosphogluconate dehydratase consititutively; in partial diploids, this constitutivity is recessive to the inducible character. It is postulated that the GntM- phenotype is due to malfunction of a negative control gene gntR, and that gntS+ specifies the activity of a gluconate uptake system.

Biological Transport, Active↗

Transcription of a lepidopteran cytochrome P450 promoter is modulated by multiple elements in its 5' UTR and repressed by 20-hydroxyecdysone.

The biochemical response to the phytochemical xanthotoxin encountered in the diet of black swallowtail larvae is the induction of P450s capable of detoxifying this and other toxic furanocoumarins. As the xenobiotic response element to xanthotoxin (XRE-xan) is necessary but not sufficient for transcription of the CYP6B1v3 gene in Sf9 cells, sequences upstream of it, such as a putative EcRE, and downstream of it, such as a putative C/EBP binding site and Inr, have been tested for their roles in regulation. Mutation of the putative EcRE has indicated that it affects basal transcription of this promoter but not repression by 20-hydroxyecdysone. Mutation of the more proximal promoter sequence, including the C/EBP and Inr, have indicated that many core promoter elements between the TATA box and translation start site modulate basal and xanthotoxin-inducible expression of this composite promoter.

5' Untranslated Regions↗