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C Pueyo

Publications and source records attributed to C Pueyo.

78 records · Page 5Linked to original sources
PubMed1979

The L-arabinose-resistance test with Salmonella typhimurium strain SV3 selects forward mutations at several ara genes.

A new assay has been described for mutagenicity testing using an L-arabinose-sensitive strain of Salmonella typhimurium. The test strain SV3 and several L-arabinose-resistant mutants selected therefrom are characterized in the present study by 3 different criteria: inhibition of growth by L-arabinose, accumulation of keto-sugars, and activities of the enzymes involved in L-arabinose catabolism. Strain SV3 (ara-531) shows high levels of inducible L-arabinose isomerase (EC 5.3.1.4) and L-ribulokinase (EC 2.7.1.16) activities, but is deficient in L-ribulose-5-phosphate 4-epimerase (EC 5.1.3.4), the enzyme encoded in Escherichia coli by gene D in the araBAD operon. Addition of L-arabinose to SV3 growing in glycerol or casamino acids stops growth. D-Glucose only partially reverses this inhibition. Reversion of the ara-531 mutation restores different levels of epimerase activity and resistance to L-arabinose. However, the great majority of the L-arabinose-resistant mutants do not utilize L-arabinose. The physiological and enzymatic properties of these L-arabinose non-utilizing mutants suggest that L-arabinose resistance is due to forward mutations in at least 3 other genes, araA, araB and araC, blocking steps prior to L-ribulose 5-phosphate accumulation.

Arabinose↗
PubMed1979

Evaluation of three metabolic activation systems by a forward mutation assay in Salmonella.

The strain SV3 of Salmonella typhimurium was used as the indicator bacterium in the intrasanguineous host-mediated mutagenicity assay. Bacterial distribution and spontaneous mutation frequency were determined after intravenous injection of SV3 into CD1 male mice. Bacteria were cleared at an exponential rate from the blood stream and recovered mainly from the liver and in smaller quantities from the lungs and kidneys. No bactericidal effect was observed during incubation within the animal, and bacterial division occurred in the liver and probably in the kidneys. The significance of an increased mutation frequency of bacteria recovered from untreated animals is discussed. Mutation induction was measured in bacteria recovered from liver, lungs and kidneys of CD1 mice and CD rats treated with dimethylnitrosamine (DMN). The sensitivity of the intrasanguineous host-mediated technique was compared with the sensitivity of the assay in vitro with microsomal preparations from each tissue and host. Activation by isolated perfused liver and lungs from CD rats was included for comparison with the results from experiments in vivo and in vitro.

Animals↗
PubMed1978

Forward mutations to arabinose resistance in Salmonella typhimurium strains: a sensitive assay for mutagenicity testing.

The forward-mutation assay using the L-arabinose-sensitive strain SV3 of Salmonella typhimurium has been calibrated against a selected set of mutagens. Strain SV3 is sensitive to chemicals causing base-pair substitutions, frameshift mutations and deletions. New strains deficient for the excision-repair system or the lipopolysaccharide barrier or both have been selected from strain SV3. The additional mutations do not affect the independence of the assay from experimental artifacts due to physiological or lethal damage or differences in plating density. The new strains are more sensitive than SV3 to certain mutagens. Techniques for using this set of strains are presented and their relative advantages discussed.

Arabinose↗
PubMed1978

A mutagen assay detecting forward mutations in an arabinose-sensitive strain of Salmonella typhimurium.

Strain SV3 of Salmonella typhimurium is sensitive to arabinose, that is, unable to grow in a medium containing arabinose plus glycerol as carbon source. Arabinose resistance is the consequence of the mutational inactivation of one of at least three different genes. The selection of arabinose-resistant mutants provides a simple and sensitive assay for the detection of weak mutagens and for refined quantitative studies of strong ones. The assay is not influenced by experimental artifacts derived from physiological or lethal effects or from differences in plating density. Such artifacts are common with other bacterial mutagen assays, including those using strains analogous to SV3. As practical examples, the assay was used with N-methyl-N'-nitro-N-nitrosoguanidine and the fungicide captafol.

Arabinose↗
PubMed1978

Genetical and biochemical characterization of QA-3 mutants and revertants in the QA gene cluster of Neurospora crassa.

The qa-3 gene, one of the four genes in the qa gene cluster, encodes quinate (shikimate) dehydrogenase (quinate: NAD oxidoreductase, ER 1.1.1.24), the first enzyme in the inducible quinic acid catabolic pathway in Neurospora crassa. Genetic analyses have localized 26 qa-3 mutants at 11 sites on the aq-3 genetic map on the basis of prototroph frequencies. Certain mutants, e.g., 336-3-10 and 336-3-3, are located at opposite ends of the qa-3 gene. Data from four-point crosses (qa-1s mutant 124 X five different qa-3 mutants in triple mutants qa-3, qa-4, qa-2) indicate the following orientation of the qa-3 gene within the qa cluster; qa-1, qa-3 mutant 336-3-10 ("left" end) qa-3 mutant 336-3-3 ("right" end), qa-4, qa-2. Ultraviolet-induced revertants have been obtained from 14 of the qa-3 mutants. The revertable mutants fall into two major classes: those that revert by changes either at the same site or at a second site within the qa-3 gene, and those that revert by unlinked suppressor mutations. The intragenic revertants can be further distinguished by quantative and/or qualitative differences in their quinate dehydrogenase activities. Some revertants with activities either equivalent to or less than wild type produce a thermostable enzyme, and others an enzyme which is thermolabile in vitro at 35 degrees. A concentration of quinic acid or shikimic acid as low as 50 micron protects the enzyme markedly from heat inactivation. The genetic organization and the orientation of the qa-3 gene are discussed with respect to its direction of transcription and to the possible localization of a promoter (initiator) region(s) within the qa gene cluster.

Alcohol Oxidoreductases↗