Promotion or retardation of the growth of adenine auxotrophs ofCandida albicans by purines, pyrimidines and nucleosides

Sarachek, Alvin

doi:10.1007/bf02046735

Cited by 29 publications

(16 citation statements)

References 20 publications

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“…The transformation data shown here support this assignment. Our data also suggest that the mutation in strain WCR-1-74 is in ADEI, an assignment made by complementation analysis (17) but in conflict with biochemical studies (20).…”

Section: Methodscontrasting

confidence: 43%

“…Strain WCR-1-74, isolated by Sarachek (20), was tested for transformation ability with pMK3 (10 jug) and pMC1 (15 ,ug DISCUSSION The results presented here establish an integrative transformation system for the pathogenic yeast C. albicans based on its cloned ADE2 gene. Because C. albicans is naturally resistant to high levels of G418 (unpublished observation), G418 resistance, used to develop transformation systems in other yeasts (4,25), can not be used as a selection for C. albicans.…”

Section: Methodsmentioning

confidence: 77%

“…Since transformation would be expected to be a rare event, this system requires a gene which functions in the host and a system for selecting individuals which have taken up and expressed this gene. The gene chosen for transformation was (12,17,20). These could arise from lesions in one of two successive steps in purine biosynthesis (6,7,24).…”

mentioning

confidence: 99%

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Integrative transformation of Candida albicans, using a cloned Candida ADE2 gene.

Kurtz¹,

Cortelyou²,

Kirsch³

1986

Mol. Cell. Biol.

163

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Candida albicans is a diploid dimorphic yeast with no known sexual cycle. The development of a DNA transformation system would greatly improve the prospects for genetic analyses of this yeast. Plasmids were isolated from a Candida Sau3A partial library which complements the ade2-1 and ade2-5 mutations in Saccharomyces cerevisiae. These plasmids contain a common region, part of which, when subcloned, produces ade2 complementation. Among the small number of auxotrophs previously isolated in C. albicans, red adenine-requiring mutants had been identified by several groups. In two of these strains, the cloned Candida DNA transformed the mutants to ADE+ at frequencies of 0.5 to 5 transformants per micrograms of DNA. In about 50% of the transformants, plasmid DNA sequences became stably integrated into the host genome and, in the several cases analyzed by Southern hybridization, the DNA was integrated at the site of the ADE2 gene in one of the chromosomal homologs.

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Section: Methodscontrasting

confidence: 43%

Section: Methodsmentioning

confidence: 77%

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Integrative transformation of Candida albicans, using a cloned Candida ADE2 gene.

Kurtz¹,

Cortelyou²,

Kirsch³

1986

Mol. Cell. Biol.

163

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“…The exceptional auxotroph (strain 758, isolated from THA) required adenine for growth and accumulated red pigment under conditions of adenine limitation. Phenotypically similar auxotrophs are known in S. cerevisiae, C. albicans, and other species (8,11,14,15). Phenotypically C These data appear also in Table 2.…”

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confidence: 54%

Parasexual genetics of Torulopsis glabrata

Whelan

Kwon-Chung

1987

J Bacteriol

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Prototrophic hybrids were generated in the asexual yeast Torulopsis glabrata by the fusion of spheroplasts derived from parent strains which bore complementing auxotrophic markers. The DNA content (per cell) of two hybrids was essentially that predicted by summing the corresponding parental values. UV irradiation of these two hybrids resulted fi the formation of sectored colonies with genetic properties consistent with their origin by either mitotic recombination or chromosomal nondisjunction.The asexual yeast Torulopsis glabrata is an opportunistic pathogen (1). In a study of fungemia in immunosuppressed patients, T. glabrata was isolated in a significant number of episodes of fungemia (26 of 136) (9). Recently, Hickey et al.(7) described a lethal infection in an immunosuppressed patient and provided a review of the literature.Previous basic studies with T. glabrata were concerned mainly with the structure of its mitochondrial genome and with the genetic basis of its killer activity against other yeasts (3,16,17). In a study of hybridization, Galeotti et al.(5) obtained T. glabrata hybrids by inducing the fusion of spheroplasts derived from parents (presumed haploid); however, those hybrids contained less DNA per cell than was predicted by summing the observed parental values of DNA per cell. Those researchers suggested, on the basis of their finding that the dose-response curves for X-ray killing of hybrids were indistinguishable from the corresponding doseresponse curves for parents, that T. glabrata lacked a recombinational pathway for repair of DNA damage.Development of a parasexual system based upon euploid T. glabrata hybrids was the object of the present study. We report here that euploid hybrids may be obtained by fusion of spheroplasts, and we provide evidence that these hybrids give rise to variants which express parental markers.MATERIALS AND METHODS Strains. T. glabrata G4 was described previously (21). The haploid Saccharomyces cerevisiae reference strain XP660-14D was provided by T. R. Manney. Candida albicans reference strain 208R1 was a derivative of clinical isolate MS24, which was described previously (18).Culture methods. The undefined complete medium YEPD agar and the defined minimal medium MIN agar were described previously (4). MIN agar was supplemented (with one or more of the compounds L-tryptophan, L-histidine, and L-methionine, to a final concentration of 40 mg/liter) as appropriate (see below). Cultures were grown at 300C, except when otherwise indicated.Induction of mutation. T. glabrata G4 was grown on YEPD agar (2 days). A sample of this confluent culture was suspended in water, spread on YEPD agar to yield approximately 600 CFU per petri dish, and exposed for 15 s to UV light delivered by a Sylvania G8T5 germicidal lamp. The dose rate at the agar surface, 32 ergs/mm2 per s, was estimated with a Blak-Ray UV meter (Ultra-Violet Products, San Gabriel, Calif.). Irradiated cultures and unirradiated * Corresponding author. controls were incubated for 2 days, and colonies were replica pl...

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“…End-product inhibition by purines has been studied in several microorganisms (Wyngaarden and Greenland, 1963;Nierlich and Magasanik, I965 a;Sarachek, 1964;Burns, 1964;Gots, 1957) Wyngaarden and Ashton, 1959)• Wyngaarden and Ashton (1959) demonstrated the regulation of purine bio synthesis via PEPP amidotransferase with high levels of purine ribonucleotides. Other inhibition sites found in clude succinoadenylate kinosynthetase by AMP and GMP, controlling production of AMP (Wyngaarden and Greenland, 1963), the regulation of GMP reductase by ATP (Magasanik, 1962), and the regulation_pf the conversion of AMP to IMP via AICAR by histidine feedback inhibition at the site con verting ATP-EPPP to "compound III" (Moyed and Magasanik, 1957)' While there are probably several sites of repres sion, only two have been discovered, both of which, were found through the use of cell-free extracts of bacteria.…”

Section: M3mentioning

confidence: 99%

The genetic control of purine biosynthesis in Staphylococcus aureus

Gabbard

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Promotion or retardation of the growth of adenine auxotrophs ofCandida albicans by purines, pyrimidines and nucleosides

Cited by 29 publications

References 20 publications

Integrative transformation of Candida albicans, using a cloned Candida ADE2 gene.

Integrative transformation of Candida albicans, using a cloned Candida ADE2 gene.

Parasexual genetics of Torulopsis glabrata

The genetic control of purine biosynthesis in Staphylococcus aureus

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