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Biomedical subjects

A K Chatterjee

Publications and source records attributed to A K Chatterjee.

At least 91 records · Page 5Linked to original sources
PubMed1985

Isolation and characterization of Tn5 insertion mutants of Erwinia chrysanthemi that are deficient in polygalacturonate catabolic enzymes oligogalacturonate lyase and 3-deoxy-D-glycero-2,5-hexodiulosonate dehydrogenase.

Mutants of Erwinia chrysanthemi EC16 deficient in the polygalacturonate catabolic enzymes oligogalacturonate lyase (Ogl-) and 3-deoxy-D-glycero-2,5-hexodiulosonate (ketodeoxyuronate) dehydrogenase (KduD-) were obtained by Tn5 mutagenesis using the R plasmid pJB4JI. Ogl- Exu+ (Exu+, D-galacturonate utilization) and KduD- Exu- strains macerated potato tuber tissue and utilized glucose, glycerol, and gluconate, but they did not utilize polygalacturonate, unsaturated digalacturonate, or saturated digalacturonate. Genetic and physical evidence indicated that the Ogl- mutants and a KduD- recombinant contained a single copy of Tn5 and that Tn5 (Kmr) was linked to the mutant phenotypes. In the Ogl+ parents, basal levels of oligogalacturonate lyase were present in glycerol-grown cells and induced levels were present with saturated or unsaturated digalacturonate, while oligogalacturonate lyase was undetectable under similar conditions in Ogl- strains. Pectate lyase, polygalacturonase, and ketodeoxyuronate dehydrogenase were induced in an Ogl- strain by 3-deoxy-D-glycero-2,5-hexodiulosonate and by the enzymatic products of unsaturated digalacturonate but not by the digalacturonates. The KduD- strains lacked the dehydrogenase activity but in the presence of the digalacturonates produced higher levels of pectate lyase, polygalacturonase, and oligogalacturonate lyase than the KduD+ parents did. In the KduD- strains, pectate lyase and oligogalacturonate lyase were induced by unsaturated digalacturonate in a "gratuitous" manner, suggesting an intracellular accumulation of the inducer(s). We conclude that an intermediate(s) of the ketodeoxyuronate pathway induces pectate lyase, polygalacturonase, oligogalacturonate lyase, and ketodeoxyuronate dehydrogenase in E. chrysanthemi.

Bacterial Proteins↗
PubMed1985

Isolation and characterization of Tn5 insertion mutants of Pseudomonas syringae pv. syringae altered in the production of the peptide phytotoxin syringotoxin.

A syringotoxin-producing strain of Pseudomonas syringae pv. syringae (B457) was subjected to Tn5 mutagenesis by the transposon vector pSUP1011. Analyses of auxotrophs obtained suggested simple random insertions of Tn5. Syringotoxin-negative mutants arose at a frequency of about 0.28%. In a Southern blot analysis, the loss of toxin production was associated with Tn5 insertions into chromosomal EcoRI fragments of about 10.5, 17.8, and 19.3 kilobases. Data from a Southern blot analysis of SstI-digested DNA from these mutants suggest that the 10.5- and 17.8-kilobase EcoRI fragments may be adjacent to or near each other. Mutants that produced only 3 to 4% wild-type toxin levels also were identified.

Bacterial Toxins↗
PubMed1985

recA is required in the induction of pectin lyase and carotovoricin in Erwinia carotovora subsp. carotovora.

Pectin lyase (PNL) and the bacteriocin carotovoricin (CTV) were induced in Erwinia carotovora subsp. carotovora 71 by the DNA-damaging agents mitomycin C, nalidixic acid, and UV light. To determine whether the recA product was involved in the expression of these damage-inducible phenotypes, we cloned the E. carotovora subsp. carotovora recA+ gene, inactivated it by Tn5 insertion, and constructed an E. carotovora subsp. carotovora recA::Tn5 strain by gene replacement via homologous recombination. The RecA- strain was more sensitive to methyl methanesulfonate, nitroquinoline oxide, and UV light than its RecA+ parent. The recA mutation did not affect the production of pectate lyase, polygalacturonase, cellulase, and protease or the ability to cause soft rot of potato tubers. With this mutant, unlike with the RecA+ parent strain, PNL and CTV were not induced by mitomycin C or detected in potato tuber tissue. The RecA+ phenotype, including the inducibility of PNL and CTV, could, however, be restored in the mutant in trans by the recA+ gene from either E. carotovora subsp. carotovora or Escherichia coli. We conclude that, in E. carotovora subsp. carotovora, the recA product is required in the induction of PNL and CTV.

Bacteriocins↗
PubMed1984

Transposon Tn5 mutagenesis in Erwinia carotovora subsp. carotovora and E. carotovora subsp. atroseptica.

In matings between Escherichia coli 2492(pJB4JI) and Erwinia carotovora subsp. carotovora Ecc71 and E. carotovora subsp. atroseptica Eca12, Kmr Gms transconjugants were obtained at high frequencies, indicating instability of the Mu-containing plasmid pJB4JI and transposition of Tn5 into the recipient genome. This was verified by Southern blot hybridization with pRZ102 DNA containing Tn5 as the 32P-labeled probe. Examination of Kmr Gms transconjugants of Ecc71 and Eca12 disclosed that a proportion (2 to 3%) were either auxotrophic or defective in catabolism of specific carbohydrates. Spontaneous prototrophic revertants were obtained for all markers with the exception of ilv, tyr, and suc. Genetic and physical data indicate that scattered insertions of Tn5 from pJb4JI into the chromosome of Ecc71 and Eca12 produced a variety of altered phenotypes due mostly to single insertions of Tn5 not accompanied by Mu DNA.

Bacteriophage mu↗
PubMed1984

Studies on certain drug-metabolizing enzymes in deoxypyridoxine-treated rats.

The effect of deoxypyridoxine on the activities of drug-metabolizing enzymes was investigated in male rats. Phenylbutazone oxidase and aminopyrine N-demethylase decreased in both liver and kidney of deoxypyridoxine-treated rats that received either an 18% or 8% casein diet. However, the magnitude of decrease in activities was more when the rats received an 8% casein diet. The NADPH oxidase activity remained unchanged following deoxypyridoxine treatment. The diminished activities of phenylbutazone oxidase and aminopyrine N-demethylase noted after deoxypyridoxine treatment were restored by pyridoxine supplementation. The decreased activities of drug-metabolizing enzymes in deoxypyridoxine treated rats were not reversed by thyroxine supplementation. It is suggested that pyridoxine in the form of pyridoxal phosphate might be involved in the regulation of drug-metabolizing activities.

Aminopyrine N-Demethylase↗
PubMed1983

Tn5-Induced Mutations in the Enterobacterial Phytopathogen Erwinia chrysanthemi.

Escherichia coli (2492/pJB4JI) matings with Erwinia chrysanthemi produced kanamycin resistant (Km) transconjugants, a majority of which were gentamicin sensitive (Gm). A small proportion (about 0.8%) of the Km Gm clones were either auxotrophic or failed to catabolize galacturonate (Gtu). The R plasmid (pJB4JI) DNA was detected in the parent E. coli strain and in a Km Gm transconjugant, but not in Km GmE. chrysanthemi strains carrying Tn5-induced mutations. In Hfr crosses, Km (Tn5) was found linked with most mutations. A majority (>95%) of prototrophic recombinants were Km, except for Leu and Arg recombinants which were 30 to 50% Km. Spontaneous revertants were obtained for all markers except car, gtu, lys, thr, and trp. Prototrophic revertants, with the exception of Met, Leu, or His clones, were Km. We conclude from both genetic and physical data that Tn5 transposed from pJB4JI into different sites on the chromosome of E. chrysanthemi.

Journal Article↗
PubMed1982

Effect of emetine treatment on subcellular nucleic acid and protein metabolism.

The subcellular RNA and protein metabolism have been studied in emetine-treated rats. Emetine treatment for a period of 10 days reduced the incorporation in vivo of [14C]-leucine into proteins of nuclear fraction of liver. This was accompanied by a reduction in the nuclear protein content. The mitochondrial fraction of heart exhibited an increase in the protein content after emetine treatment. This was, however, not accompanied by an increased rate of incorporation into its protein. On the other hand, the nuclear fraction of heart of emetine-treated rats showed a diminution in the rate of incorporation into its protein without having been reduced in protein content. The mitochondrial and microsomal RNA of liver and the microsomal RNA of heart responded to emetine treatment by showing increased levels. The liver RNase activity was reduced after emetine treatment, while the heart RNase remained independent of the treatment. It has been suggested that both synthesis and breakdown of liver proteins are reduced by emetine treatment. The synthesis of liver nuclear proteins was supposedly affected more than was its breakdown by emetine. The synthesis of rapidly turningover proteins of heart nuclear fraction and the catabolism of slowly turningover proteins of its mitochondrial fraction are thought to be reduced by the emetine treatment.

Animals↗
PubMed1982

Studies on the possible mechanism of hydrazine action on ascorbic acid-metabolising enzymes.

The activities of enzymes associated with the synthesis and degradation of L-ascorbic acid were studied in hydrazine-treated rats supplemented with pyridoxine. The effects of hydrazine in vitro were also examined on these enzymes. The activity of liver D-glucuronoreductase was reduced in hydrazine-treated rats even after receiving pyridoxine in excess. But, the decreased activities of liver and kidney dehydroascorbatases in hydrazine-treated rats were reversed by pyridoxine supplementation. Hydrazine in vitro could not inhibit the activity of liver D-glucuronoreductase or L-gulonooxidase. The drug was also unable to inhibit in vitro the activity of liver and kidney dehydroascorbatases. Studies with dialyzed liver homogenates showed that the diminished activity of D-glucuronoreductase in the liver of hydrazine-treated rats was maintained even after dialyzing the tissue preparations. In contrast, the reduced activity of dehydroascorbatase in the liver of hydrazine-treated rats was abolished following dialysis of the liver preparations. It has been suggested that the diminished activity of liver D-glucuronoreductase after hydrazine treatment might arise from the reduced synthesis of enzyme protein, while the reduction in the activity of dehydroascorbatase following hydrazine treatment could be ascribed to depletion in vivo of pyridoxal phosphate and/or to the involvement of some dialyzable factors.

Animals↗
PubMed1980

Possible influence of gonadotrophins on formation in vivo of pyridoxal phosphate.

FSH administered to normal rats increased the activity of pyridoxine phosphate oxidase of both liver and kidney and, consequently, pyridoxal phosphate levels in these tissues were elevated. LH administration, on the other hand, decreased the activity of pyridoxine phosphate oxidase, resulting in diminished pyridoxal phosphate level in the tissues. The stimulatory effect of FSH on the activity of liver and kidney pyridoxine phosphate oxidase was not observed in castrated-adrenalectomised rats unless supplemented with cortisone and testosterone, respectively. Puromycin treatment prevented the FSH-induced rise in the activity of liver and kidney pyridoxine phosphate oxidases. It is suggested that FSH stimulates the activity of liver and kidney pyridoxine phosphate oxidase by increasing the synthesis of apoproteins of the enzyme, and the effect of FSH on liver is dependent on the presence of adrenal corticoids while the presence of testosterone is a prerequisite for the FSH to have its effect on kidney pyridoxine phosphate oxidase.

Adrenalectomy↗
PubMed1980

Acceptance by Erwinia spp. of R plasmid R68.45 and its ability to mobilize the chromosome of Erwinia chrysanthemi.

R plasmid R68.45 was transferred in broth matings from Escherichia coli to strains of Erwinia amylovora, E. carotovora subsp. atroseptica, E. chrysanthemi, and E. herbicola (Enterobacter agglomerans); the frequency of transfer ranged from 2 x 10(-8) to 5 x 10(-4) per input donor cell depending on the bacterial species. The drug resistance markers tet(+), amp(+), and kan(+) were stable in these Erwinia species. Transconjugants of Erwinia spp., but not of the wild-type parent Erwinia strains, acquired levels of antibiotic resistance (tetracycline, 50 mug/ml; ampicillin, 200 mug/ml; kanamycin 200 mug/ml) similar to those of the donor R68.45-bearing strain of Escherichia coli. Erwinia transconjugants (with one exception of E. carotovora subsp. atroseptica) were donors of the antibiotic resistance markers; the frequency of transfer was consistently higher with an E. coli strain than with Erwinia spp. as recipients, and when matings were done on a solid surface (membranes) rather than in liquid. Transfer of chromosomal markers ade(+), gal(+), gtu(+) (utilization of galacturonate), his(+), leu(+), lys(+), thr(+), and trp(+) occurred in crosses between E. chrysanthemi strains harboring R68.45 and appropriate recipient strains; the frequency of transfer ranged from 9.0 x 10(-8) to 2.0 x 10(-6) depending on the selective marker. Analysis of the coinheritance of unselected markers among various classes of recombinants revealed linkage between thr-leu-lys-ade and between trp and his, thus confirming earlier findings with the Hfr-type donor cells. Since R68.45 mobilized an array of chromosomal markers in the wild-type as well as genetically marked strains of E. chrysanthemi, the system, used in conjunction with the existing Hfr strains, should provide a useful tool to study the genetics of plant pathogenicity of this bacterial species. In contrast to E. chrysanthemi, R68.45 did not mobilize chromosomal markers ilv(+), his(+), rbs(+), ser(+), and thr(+) in E. amylovora EA178.

Ampicillin↗
PubMed1980

Generalized transduction in the enterobacterial phytopathogen Erwinia chrysanthemi.

Bacteriophages induced by mitomycin treatment of Erwinia chrysanthemi KS612 produced plaques on lawns of E. chrysanthemi EC183 and KS605. Bacteriophage Erch-12, purified from one such plaque, transferred an array of chromosomal genes (arg, leu, his, ser, thr, trp, ura) to appropriate recipient strains derived from E. chrysanthemi EC 183. Recombinants were formed in the absence of cellular contact between donor and recipient bacteria and in the presence of deoxyribonuclease. Ultraviolet irradiation of the bacteriophage stimulated transductional frequency. Linkage was detected in two-factor crosses between the loci thr and ser and between rif and ade; several closely linked mutations in ser were mapped with respect to thr.

Amino Acids↗