Regulation of β-Galactosidase Synthesis in Wild Type and in a Succinate-Resistant Mutant of Rhizobium meliloti

1985 ◽  
Vol 40 (3-4) ◽  
pp. 170-175
Author(s):  
A. P. Singh ◽  
J. B. Singh

The synthesis of β-galactosidase in Rhizobium meliloti WU60 was found to be inducible by lactose and its non-metabolizable analogue. isopropyl-β-ᴅ-thiogalactoside (IPTG). In contrast to Escherichia coli, galactose and melibiose were very weak inducers of this enzyme in R. meliloti. The maximum level of β-galactosidase in this bacterium is 2% of that in fully induced E. coli. In addition to glucose, the induced synthesis of this enzyme in R. meliloti was repressed by galactose, glycerol, and succinate. In comparison to E. coli, addition of cyclic AMP to the growth medium of R. meliloti did not alleviate the repressive effect of the above compounds on β- galactosidase synthesis. High concentrations of sodium succinate (100 mᴍ) were inhibitory to the growth of R. meliloti. Spontaneous succinate-resistant mutants could be isolated at low frequency. In contrast to the wild type parent, in a succinate-resistant mutant, the synthesis of β-galactosidase was not repressed by succinate.

2002 ◽  
Vol 48 (5) ◽  
pp. 458-462 ◽  
Author(s):  
Chengru Zhu ◽  
Musangu Ngeleka ◽  
Andrew A Potter ◽  
Brenda J Allan

The Fur (ferric uptake regulator) protein is a master regulator of iron metabolism in gram-negative bacteria. In the present study, the effect of a partial deletion of the fur gene on the acid-tolerance response and in vivo virulence of avian Escherichia coli was examined. The fur mutant was unable to trigger the acid-tolerance response as observed in the wild-type parent strain. However, the mutant was as virulent as the wild-type parent strain when tested in 1-day-old chickens by subcutaneous inoculation. These data indicate that the fur gene is involved in the acid-tolerance response but not involved in the virulence of E. coli, as detected by the ability to cause septicemia in our experimental infection.Key words: E. coli, fur, acid-tolerance response.


1975 ◽  
Vol 75 (2) ◽  
pp. 275-292 ◽  
Author(s):  
H. Williams Smith ◽  
J. F. Tucker

SUMMARYChickens in groups of 40 were infected orally with a nalidixic acid-resistant mutant of Salmonella typhimurium and then fed continuously on diets containing ampicillin, chloramphenicol, furazolidone, neomycin, oxytetracycline, polymixin, spectinomycin, streptomycin or a mixture of trimethoprim and sulphadiazine. The amount of S. typhimurium excreted in their faeces was estimated at intervals by culture on brilliant green agar containing sodium nalidixate, both direct and after enrichment in selenite broth; the amount of Escherichia coli excreted was estimated by culture on MacConkey agar. The feeding of diets containing 500 mg./kg. of ampicillin, furazolidone, neomycin, polymixin, spectinomycin or streptomycin or 100 mg./kg. of trimethoprim and 500 mg./kg. of sulphadiazine for 46 days reduced to a varying degree the amount of S. typhimurium and E. coli excreted, the greatest reduction in S. typhimurium being brought about by the last treatment. The effect was less obvious when the concentration of the antibiotics in the food was decreased fivefold. An important reason for the very limited effect of some of the antibiotics was the emergence of antibiotic-resistant populations of S. typhimurium and E. coli. High concentrations of antibiotic-resistant organisms also arose in the faeces of the chickens fed diets containing tetracyclines and chloramphenicol, treatments which had no apparent effect on the amount of S. typhimurium and E. coli excreted. Much of the antibiotic resistance encountered was determined by R factors, a particular R factor usually being found in the E. coli populations of individual chickens before it was found in their S. typhimurium populations. No S. typhimurium or E. coli were isolated that possessed R factors determining resistance to polymixin, furazolidone or trimethoprim. No S. typhimurium or E. coli were isolated that were polymixin-resistant and no S. typhimurium that were furazolidone-resistant. The few trimethoprim-resistant S. typhimurium isolated were thymine-dependent.The feeding of diets containing the higher concentrations of trimethoprim/sulphadiazine, neomycin, furazolidone or ampicillin for 9 days reduced the amount of S. typhimurium excreted. After the withdrawal of these diets, the amount of S. typhimurium excreted increased to the numbers found in chickens given ordinary diets throughout; the chickens that had been given trimethoprim/sulphadiazine or furazolidone did not remain faecal excreters of S. typhimurium longer than the chickens that had been given ordinary diets. Similar results were obtained with trimethoprim/sulphadiazine when the start of the 9-day treatment period was delayed for an extra 9 days or when it was extended to 18 days.


2000 ◽  
Vol 66 (11) ◽  
pp. 4764-4771 ◽  
Author(s):  
Takane Katayama ◽  
Hideyuki Suzuki ◽  
Takashi Koyanagi ◽  
Hidehiko Kumagai

ABSTRACT Tyrosine phenol-lyase (Tpl), which can synthesize 3,4-dihydroxyphenylalanine from pyruvate, ammonia, and catechol, is a tyrosine-inducible enzyme. Previous studies demonstrated that thetpl promoter of Erwinia herbicola is activated by the TyrR protein of Escherichia coli. In an attempt to create a high-Tpl-expressing strain, we cloned the tyrRgene of E. herbicola and then randomly mutagenized it. Mutant TyrR proteins with enhanced ability to activate tplwere screened for by use of the lac reporter system inE. coli. The most increased transcription oftpl was observed for the strain with the mutanttyrR allele involving amino acid substitutions of alanine, cysteine, and glycine for valine-67, tyrosine-72, and glutamate-201, respectively. A tyrR-deficient derivative of E. herbicola was constructed and transformed with a plasmid carrying the mutant tyrR allele (V67A Y72C E201G substitutions). The resultant strain expressed Tpl without the addition of tyrosine to the medium and produced as much of it as was produced by the wild-type strain grown under tyrosine-induced conditions. The regulatory properties of the mutant TyrRV67A, TyrRY72C, TyrRE201G, and TyrRV67A Y72C E201G proteins were examined in vivo. Interestingly, as opposed to the wild-type TyrR protein, the mutant TyrRV67A protein had a repressive effect on the tyrP promoter in the presence of phenylalanine as the coeffector.


1993 ◽  
Vol 264 (2) ◽  
pp. C464-C470 ◽  
Author(s):  
S. Fukayama ◽  
A. H. Tashjian ◽  
F. R. Bringhurst

We have used wild-type and adenosine 3',5'-cyclic monophosphate (cAMP)-resistant mutant osteoblast-like SaOS-2 cells to investigate the role of protein kinase A (PKA) in the regulation of cytosolic free Ca2+ concentration ([Ca2+]i). Basal levels of [Ca2+]i were the same in wild-type (127 +/- 6.1 nM) and transfected (117 +/- 6.8 nM) SaOS-2 cells, although 45Ca2+ efflux was slower in the transfected cells. In wild-type cells, thapsigargin (TG, > or = 200 nM), an inhibitor of the Ca(2+)-ATPase activity of the endoplasmic reticulum, acutely increased [Ca2+]i (by up to 2-fold), which then returned promptly to basal [Ca2+]i. In cAMP-resistant cells, TG elicited a significantly greater acute rise in [Ca2+]i, which then decayed to an elevated plateau level. In mutant cells, high concentrations of dibutyryladenosine 3',5'-cyclic monophosphate, which overcome the PKA blockade, restored the changes in [Ca2+]i to the wild-type pattern. In cAMP-resistant, TG-blocked cells, ionomycin (or alpha-thrombin) induced a further elevation in [Ca2+]i, which then declined rapidly to the original basal level. We conclude that basal PKA activity is involved actively in regulation of [Ca2+]i in SaOS-2 cells by promoting Ca2+ efflux from the cell and, possibly, by inhibiting Ca2+ release from or stimulating net Ca2+ sequestration into the ER. We have also obtained evidence for an alternate Ca(2+)-triggered Ca2+ reuptake mechanism in SaOS-2 cells that is not dependent on either Ca(2+)-ATPase or PKA.


2015 ◽  
Vol 25 (2-3) ◽  
pp. 209-225 ◽  
Author(s):  
Sarah L. Sutrina ◽  
Kia Daniel ◽  
Michael Lewis ◽  
Naomi T. Charles ◽  
Cherysa K.E. Anselm ◽  
...  

We established that <i>Escherichia coli </i>strain 15 (ATCC 9723) produces both curli and cellulose, and forms robust biofilms. Since this strain is wild type with respect to the phosphoenolpyruvate:sugar phosphotransferase system (PTS), it is an ideal strain in which to investigate the effects of the PTS on the biofilm growth of <i>E. coli</i>. We began by looking into the effects of PTS and non-PTS sugars on the biofilm growth of this strain. All the sugars tested tended to activate biofilm growth at low concentrations but to inhibit biofilm growth at high concentrations. Acidification of the medium was an inhibitory factor in the absence of buffer, but buffering to prevent a pH drop did not prevent the inhibitory effects of the sugars. The concentration at which inhibition set in varied from sugar to sugar. For most sugars, cyclic (c)AMP counteracted the inhibition at the lowest inhibitory concentrations but became ineffective at higher concentrations. Our results suggest that cAMP-dependent catabolite repression, which is mediated by the PTS in <i>E. coli</i>, plays a role in the regulation of biofilm growth in response to sugars. cAMP-independent processes, possibly including Cra, also appear to be involved, in addition to pH effects.


1996 ◽  
Vol 40 (5) ◽  
pp. 1266-1269 ◽  
Author(s):  
J D Goldman ◽  
D G White ◽  
S B Levy

The multiple antibiotic resistance (mar) locus in Escherichia coli consists of two divergently expressed operons (marC and marRAB), both of which contribute to the Mar phenotype. Overexpression of the marRAB operon protected E. coli against rapid cell killing by fluoroquinolones. Inactivation of the operon in mar mutants restored a wild-type bactericidal susceptibility. Both operons of the locus were required for protection from the quinolone-mediated bactericidal activity in mar locus deletion mutants. The effect was lost at high concentrations of fluoroquinolones, unlike the case for the previously described genes hipA and hipQ. The inducible mar locus appears to specify a novel antibactericidal mechanism which may play a role in the emergence of fluoroquinolone-resistant clinical E. coli isolates.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hidetada Hirakawa ◽  
Kazutomo Suzue ◽  
Ayako Takita ◽  
Chikako Awazu ◽  
Jun Kurushima ◽  
...  

Abstract The Tol-Pal system is a protein complex that is highly conserved in many gram-negative bacteria. We show here that the Tol-Pal system is associated with the enteric pathogenesis of enterohemorrhagic E. coli (EHEC). Deletion of tolB, which is required for the Tol-Pal system decreased motility, secretion of the Type III secretion system proteins EspA/B, and the ability of bacteria to adhere to and to form attaching and effacing (A/E) lesions in host cells, but the expression level of LEE genes, including espA/B that encode Type III secretion system proteins were not affected. The Citrobacter rodentium, tolB mutant, that is traditionally used to estimate Type III secretion system associated virulence in mice did not cause lethality in mice while it induced anti-bacterial immunity. We also found that the pal mutant, which lacks activity of the Tol-Pal system, exhibited lower motility and EspA/B secretion than the wild-type parent. These combined results indicate that the Tol-Pal system contributes to the virulence of EHEC associated with the Type III secretion system and flagellar activity for infection at enteric sites. This finding provides evidence that the Tol-Pal system may be an effective target for the treatment of infectious diseases caused by pathogenic E. coli.


1992 ◽  
Vol 38 (6) ◽  
pp. 534-540 ◽  
Author(s):  
Kai-Uwe Riedel ◽  
Bernd Masepohl ◽  
Werner Klipp ◽  
Alfred Pühler

To analyze the overexpression of the Rhizobium meliloti fdxN gene in Escherichia coli, different translational and transcriptional fusions were constructed. The translational signals of R. meliloti fdxN were recognized in E. coli as demonstrated by the use of in-frame lac fusions. Translational fusions consisting of the lacZ or the lpp gene fused in frame to the 3′ end of the entire fdxN gene were expressed at high levels in E. coli. In contrast, the wild-type R. meliloti FdxN protein without a C-terminal fusion could only be detected using the very sensitive T7 promoter–polymerase system and not in immunoblots with antibodies against an FdxN–LacZ hybrid protein. Evidently, translational fusions to the 3′ end of fdxN had a stabilizing effect on the expression of the fdxN gene. A constitutively expressed transcriptional fdxN fusion, which did not mediate detectable amounts of FdxN protein either in E. coli or in free-living R. meliloti cells, complemented the Fix− phenotype of an R. meliloti fdxN::[Tc] mutant strain to wild-type levels. Therefore, either low amounts of the wild-type FdxN protein are sufficient for symbiotic nitrogen fixation or there are stabilizing factors, which are present only in R. meliloti bacteroids but not in free-living R. meliloti cells. Fusion proteins consisting of FdxN and LacZ or a partial Lpp protein restored the Fix− phenotype of an R. meliloti fdxN mutant to 3 and 11%, respectively, indicating that a C-terminal fusion did not completely abolish the function of FdxN. Key words: overexpression, protein stability, immunoblot, complementation.


1984 ◽  
Vol 30 (9) ◽  
pp. 1179-1183 ◽  
Author(s):  
E. S. P. Bromfield

Variation in nodulation preference within and between cultivars of Medicago sativa for Rhizobium strains was assessed under axenic conditions using inocula consisting of paired strains of R. meliloti which could be recognized by distinctive colony morphology or antibiotic resistance. The largest variability in host preference for Rhizobium strains was among plants within cultivars and not between cultivars. The implications of such variation are discussed in the context of possible enhancement of symbiotic nitrogen fixation. An isolate of R. meliloti which had been marked with antibiotic resistance and cured of a cryptic plasmid was significantly less successful in nodulation than its wild-type parent strain in all inoculated combinations. The interaction of inoculum and cultivar on yield indicated differential symbiotic effectiveness of strains of R. meliloti on cultivars of M. sativa.


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