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1.
Nucleotide sequences from messenger RNA transcribed from the operator-proximal portion of the tryptophan operon of Escherichia coli 总被引:4,自引:0,他引:4
32P-labeled messenger RNA transcribed in vivo from the operator-proximal portion of the tryptophan operon of Escherichia coli was purified by DNA/RNA hybridization. The mRNA preparations obtained were subjected to polyaerylaamide gel electrophoresis, and a number of discrete labeled bands were detected. Characterization of the labeled bands and of purified, unbanded mRNA preparations, by partial sequence analysis of the oligonucleotides obtained following T1 and pancreatic RNase digestion, revealed that the bands represented discrete segments of the trp mRNA molecule. This observation suggests that endonucleolytic cleavage occurs in vivo at specific sites in the mRNA molecule. 相似文献
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Different half-lives of messenger RNA corresponding to different segments of the tryptophan operon of Escherichia coli 总被引:5,自引:0,他引:5
In genetically derepressed strains (trpR−) of Escherichia coli which are growing exponentially, messenger RNA regions corresponding to different segments of the trp operon are labeled with different kinetics, suggesting that operator-proximal and distal regions of trp-mRNA have different half-lives. This conclusion was confirmed by direct measurement of trp-mRNA decay; the half-lives for different mRNA regions at 30 °C were found to be 60 seconds for trpE-mRNA, 75 seconds for trpDC-mRNA, and 95 to 115 seconds for trpBA-mRNA. Deletions of genetic segments within the operator-proximal region of the operon reduce the half-life of trp BA-mRNA. Large deletions which place the BA region near the operator reduce the half-life of trpBA-mRNA to values similar to that of trpE-mRNA in the parental strain. Therefore location in the message rather than primary structure appears to determine the half-life of each mRNA region. Several of the internal deletions have a polar effect on the synthesis of the trpB and trpA polypeptides. However, the reduction in trpBA-mRNA half-life does not appear to be due to polarity because trpBA-mRNA half-life is reduced to the same value in three deletion mutants in which there is a sevenfold difference in polarity. These results are compatible with a model of trp-mRNA degradation in which the initial degradative event occurs near the 5′ end of the mRNA molecule and is followed by over-all degradation in the 3′ direction, with random or non-random delays causing an increase in half-life of about 10% per 1000 nucleotides mRNA. Our findings are not compatible with a model of normal degradation in which the entire mRNA molecule is the target for the initial degradative event. 相似文献
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Promotion, termination, and anti-termination in the rpsU-dnaG-rpoD macromolecular synthesis operon of E. coli K-12 总被引:18,自引:0,他引:18
Summary The regulatory regions for the rpsU-dnaG-rpoD macromolecular synthesis operon have been fused to a structural gene whose product is readily assayed (the Cmr structural gene coding for chloramphenicol acetyl transferase, CAT). The promoters (P1, P2, P3, Pa, Pb, Phs) for the macromolecular synthesis operon have different strengths as shown by their relative abilities to drive expression of the CAT gene. Promoter occlusion by P1 can be demonstrated within this operon. Regions 5kb upstream have a profound effect on operon gene expression. There is a thermoinducible promoter located within the dnaG structural gene. One of the macromolecular synthesis operon promoters is under lexA control. Although the operon structure allows coordinate expression of rpsU, dnaG and rpoD these additional features suggest that expression of individual genes can be independently regulated in response to altered growth conditions.Abbreviations Apr
ampicillin resistance
- CAT
chloramphenicol acetyl transferase
- Cmr
chloramphenicol resistance
- kb
kilobase pair
- orf
open reading frame
- P
promoter
- T
terminator
- Tcr
tetracycline resistance 相似文献
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Metabolism of messenger RNA from the gal operon of Escherichia coli 总被引:13,自引:0,他引:13
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Internal promoter of the tryptophan operon of Escherichia coli is located in a structural gene 总被引:22,自引:0,他引:22
The constitutive low-efficiency promoter site (P2) near the middle of the tryptophan operon of Escherichia coli has been mapped by analysis of short deletions internal to the trp operon. Comparison of deletions which remove this internal promoter with those which retain it show that P2 is located within trpD, the region coding for phosphoribosyl anthranilate transferase. P2 maps near the operator-distal end of trpD, on the operator-proximal side of two trpD point mutants. Comparisons of strains with and without the P2 site indicate that initiations at this promoter are responsible for synthesis of 80% of the trpC, trpB and trp A polypeptides present in repressed cells. 相似文献
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Mercury pollution has emerged as a major problem in industrialized zones and presents a serious threat to environment and
health of local communities. Effectiveness and wide distribution of mer operon by horizontal and vertical gene transfer in its various forms among large community of microbe reflect importance
and compatibility of this mechanism in nature. This review specifically describes mer operon and its generic molecular mechanism with reference to the central role played by merA gene and its related gene products. The combinatorial action of merA and merB together maintains broad spectrum mercury detoxification system for substantial detoxification of mercurial compounds. Feasibility
of mer operon to coexist with antibiotic resistance gene (amp
r
, kan
r
, tet
r
) clusters enables extensive adaptation of bacterial species to adverse environment. Flexibility of the mer genes to exist as intricate part of chromosome, plasmids, transposons, and integrons enables high distribution of these genes
in wider microbial gene pool. Unique ability of this system to manipulate oligodynamic property of mercurial compounds for
volatilization of mercuric ions (Hg2+) makes it possible for a wide range of microbes to tolerate mercury-mediated toxicity. 相似文献
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CLIFF E. SINGER GERALD R. SMITH RICCARDO CORTESE BRUCE N. AMES 《Nature: New biology》1972,238(81):72-74
TRANSFER RNA has been implicated in the regulation of a number of amino-acid biosynthetic operons1–4. Histidyl-tRNAHis has been shown to be involved in regulation of the histidine operon by analysis of six genes (hisO, hisR, hisS, hisT, hisU, hisW), mutation of which causes derepression of the enzymes of the histidine biosynthetic pathway in Salmonella typhimurium5–7. A class of derepressed mutants (hisR) has only about 55% as much tRNAHis as the wild type4 and in the one example sequenced, contains tRNAHIS with a structure identical to that of the wild type8. Studies of mutants of the gene for histidyl-tRNA synthetase (hisS) indicated that the derepressed phenotype was associated with defects in the charging of tRNAHISin vitro2. The amounts of charged and uncharged tRNAHis present in vivo during physiological derepression of the wild type and in the six classes of regulatory mutants, have been determined9. This work has shown that repression of the histidine operon is correlated directly with the concentration of charged histidyl-tRNAHisin vivo and not with the ratio of charged to uncharged or the absolute amount of uncharged tRNAHis. The derepression observed in mutants, of hisS (the gene for histidyl-tRNA synthetase), hisR (the presumed structural gene for the single species of tRNAHis) and hisU and hisW (genes presumably involved in tRNA modification) may be explained by the lower cellular concentration of charged tRNAHis which these mutants contain. 相似文献
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The hisG gene is the most operator-proximal structural gene of the histidine operon; it encodes the feedback-inhibitable first enzyme of the biosynthetic pathway. Previously, hisG mutants were mapped into seven intervals defined by the availble deletion mutations having endpoints in the hisG gene. The map has been refined using over 60 new deletion mutants. The new map divides the gene into 40 deletion intervals, which average approximately 30 base pairs in length. The map has been used to analyze the distribution of insertion sites for the transposable element Tn10 and has permitted conclusions on the diistribution of duplication endpoints. The map promises to be useful in analysis of his regulation and, more particularly, in the determination of the possible role of the hisG enzyme in this mechanism. 相似文献
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Summary Four genes, nagR, A, B and E, clustered in the nag locus of Escherichia coli K12 and Klebsiella pneumoniae, were cloned and physically mapped, and the corresponding gene products involved in amino sugar metabolism identified. Expression of the nag genes was also analysed using a series of lacZ fusions. In both bacteria, the genes are arranged in two divergent operons and controlled by a common NagR repressor. The corresponding gene nagR was found to map in the first operon together with the promoter proximal gene nagB, encoding the enzyme d-glucosamine isomerase (deaminase) (NagB) and the middle gene nagA, coding for N-acetyl-glucosamine deacetylase (NagA). Polar mutations in nagB and nagA prevent the efficient expression of nagR and cause constitutive expression of all nag genes. This includes the gene nagE encoding Enzyme IINag of the phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS), encoded in the second divergently transcribed operon. No further gene is found in this operon which in both organisms is directly adjacent to the gene glnS. It is interesting that the NagR repressor also affects the mannose PTS (genes manX, Y, Z), the second transport system involved in amino sugar uptake and phosphorylation. 相似文献
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Primary bile acids are synthesized from cholesterol in the liver, conjugated to either glycine or taurine and secreted into bile. Bile salts undergo enterohepatic circulation several times each day. During this process, they are biotransformed into a variety of metabolites by gut bacteria. The major biotransformation is the 7α-dehydroxylation of cholic acid and chenodeoxycholic acid yielding deoxycholic acid and lithocholic acid, respectively. 7α-Dehydroxylation is a multi-step pathway. The genes encoding enzymes in this pathway have been identified in two species of “high” activity strains of clostridia. Here, we report the isolation and characterization of a bile acid inducible (bai) operon in Clostridium hylemonae, a “low” activity 7α-dehydroxylating strain. The gene organization and sequence of the baiBCDEFGHI operon was highly conserved between C. hylemonae and “high” activity strains. Surprisingly, the baiA gene was missing from the bai operon of C. hylemonae. The baiA gene was isolated using PCR and degenerate oligonucleotide primers. The mRNA start site for the large bai operon was determined and shown to be only 11 bp from the initiation codon of the first gene. It was also discovered that allocholic acid (5α) induced the bai operon and stimulated the conversion of [24-14C] cholic acid to [24-14C] allodeoxycholic acid in cultures of C. scindens and C. hylemonae allodeoxycholic acid. Finally, it was discovered that the addition of testosterone to the growth medium markedly increased 7α-dehydroxylation of cholic acid in Clostridium scindens and C. hylemonae. We hypothesize that testosterone may be a gratuitous inducer of genes involved in the reductive arm of the bile acid 7α-dehydroxylation pathway. 相似文献
19.
Glycogen in Bacillus subtilis: molecular characterization of an operon encoding enzymes involved in glycogen biosynthesis and degradation 总被引:7,自引:1,他引:7
Although it has never been reported that Bacillus subtilis is capable of accumulating glycogen, we have isolated a region from the chromosome of B. subtilis containing a glycogen operon. The operon is located directly downstream from trnB, which maps at 275 on the B. subtilis chromosome, it encodes five poly-peptides with extensive similarity to enzymes involved in glycogen and starch metabolism in both prokaryotes and eukaryotes. The operon is presumably expressed by an EσE-controlled promoter, which was previously identified downstream from trnB. We have observed glycogen biosynthesis in B. subtilis exclusively on media containing carbon sources that allow efficient sporulation. Sporulation-independent synthesis of glycogen occurred after integration of an EσA controlled promoter upstream of the operon. 相似文献
20.
The sodium -pumping NADH: ubiquinone oxidoreductase (Na+-NQR) is the main ion pump and the primary entry site for electrons into the respiratory chain of many different types of pathogenic bacteria. This enzymatic complex creates a transmembrane gradient of sodium that is used by the cell to sustain ionic homeostasis, nutrient transport, ATP synthesis, flagellum rotation and other essential processes. Comparative genomics data demonstrate that the nqr operon, which encodes all Na+-NQR subunits, is found in a large variety of bacterial lineages with different habitats and metabolic strategies. Here we studied the distribution, origin and evolution of this enzymatic complex. The molecular phylogenetic analyses and the organizations of the nqr operon indicate that Na+-NQR evolved within the Chlorobi/Bacteroidetes group, after the duplication and subsequent neofunctionalization of the operon that encodes the homolog RNF complex. Subsequently, the nqr operon dispersed through multiple horizontal transfer events to other bacterial lineages such as Chlamydiae, Planctomyces and α, β, γ and δ -proteobacteria. Considering the biochemical properties of the Na+-NQR complex and its physiological role in different bacteria, we propose a detailed scenario to explain the molecular mechanisms that gave rise to its novel redox- dependent sodium -pumping activity. Our model postulates that the evolution of the Na+-NQR complex involved a functional divergence from its RNF homolog, following the duplication of the rnf operon, the loss of the rnfB gene and the recruitment of the reductase subunit of an aromatic monooxygenase. 相似文献