Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans

Swoboda, Rolf; Bertram, Gwyneth; Colthurst, David R.; Tuite, Mick F.; Gow, Neil A. R.; Gooday, Graham W.; Brown, Alistair J. P.

doi:10.1099/00221287-140-10-2611

Cited by 24 publications

(38 citation statements)

References 31 publications

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“…The Ura-strain C. albicans SGY243 was a gift from R. Kelly (Kelly et al, 1987). NGY2 and NGY4 are heterozygous Achs2 null mutants with one or two disrupted copies of CaCHS2 and NGY 10 is a homozygous Achs2 null mutant (Gow et al, 1994). All were constructed from the parental Ura-strain SGY243 (Table 1).…”

Section: Strains and Mutantsmentioning

confidence: 99%

“…albicans cells were harvested at the various time-points by centrifugation and cell pellets were stored at -70 "C. Total RNA was extracted from the cells and Northern blots were prepared by the method of Hube et al (1994). N o suitable internal loading standard has been identified for Northern analysis of C. albicans mRNA during germ tube formation (Delbruck & Ernst, 1993;Swoboda et al, 1994;Gow et al, 1995). Therefore, equal loading was confirmed by examining the density of ethidium-bromide-stained rRNA bands.…”

Section: Strains and Mutantsmentioning

confidence: 99%

“…One discrepancy between the different studies has been the chitin content of the Achs2 null mutant. Gow et al (1994) reported a 40 '/o reduction in the chitin content of the hyphal form of a A h 2 null mutant whereas Mio et al (1996) found the chitin content to be similar to that of the parental strain. We address this discrepancy and report here that chitin contents determined by different methods yield markedly different results that can account for the apparent discrepancies in the literature.…”

Section: Introductionmentioning

confidence: 99%

“…Characterization of the role of Candida chitin synthase isoenzymes has employed the Ura-blaster technique to disrupt the CaCHS2 (Gow et al, 1994) and the CaCHS3 (Bulawa et al, 1995) genes. Neither of these genes was found to be essential and both null mutants were able to produce hyphae.…”

Section: Introductionmentioning

confidence: 99%

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Regulation of chitin synthesis during dimorphic growth of Candida albicans

et al. 1998

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Candida albicans has three genes encoding chitin synthase enzymes. In wildtype strains, the expression of CHS2 and CHS3 peaked 1-2 h after the induction of hyphal growth, whilst mRNA levels in a non-germinative strain, CA2, remained low under the same conditions. CHSl gene expression did not peak during germ tube formation but remained a t low levels in both yeast and hyphal growth. The pattern of gene expression did not predict the changes in measured chitin synthase activities or changes in chitin content during dimorphic transition. Chitin synthase activity increased steadily, and did not peak shortly after germ tube induction, and activity profiles were similar in germ-tube-competent and germ-tube-negative strains. The phenotype of a Achs2 null mutant suggested that CHS2 encoded the major enzyme activity in vitro and was largely responsible for elevated chitin synthase activities in microsomal preparations from hyphal cells compared to yeast cells. However, CaChs3p was responsible for synthesis of most chitin in both yeast and hyphae. Three independent chitin assays gave markedly different estimates of the relative chitin content of yeast and hyphae and wild-type and chs mutants. Only one of the methods gave a significantly higher chitin content for hyphal compared to yeast cell walls and a lower chitin content for hyphae of the Achs2 null mutant compared to the parental strain.

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Section: Strains and Mutantsmentioning

confidence: 99%

Section: Strains and Mutantsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Regulation of chitin synthesis during dimorphic growth of Candida albicans

et al. 1998

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“…albicans are dimorphic fungi, 3,4) and the pathogenicity of hyphal cells is stronger than that of yeast cells at the infected site. [12][13][14] The cAMP signal pathway is a regulation system of the transformation.…”

Section: Discussionmentioning

confidence: 99%

Hyphae Formation of Candida albicans Is Regulated by Polyamines

Ueno

Fukumatsu

Ogasawara

et al. 2004

Biological & Pharmaceutical Bulletin

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Candida albicans are opportunistic fungi which generally exist in the oral cavity, skin, vagina and intestinal organs, 1,2) and are dimorphic fungi which undergo a transition from the yeast form to the hyphal form depending on the growth conditions.3,4) They grow in yeast form at membranes and the surface of the skin, but in hyphae form at the deep-seated mycosis.Temperature, pH and Ca 2ϩ are known as external factors of the transformation.5-7) The hyphae transformation is induced by the activation of the MAP kinase and cAMP pathways, which are controlled by RAS protein. 8,9) Polyamines are synthesized from ornithine by ornithine decarboxylase (ODC) and known as inducers of adenylate cyclase. 10)In this report, we discovered that polyamines participated in the transformation of C. albicans, and the activation of adenylate cyclase was related to the transformation. MATERIALS AND METHODSOrganism and Growth Conditions C. albicans NIH A-207 strain was routinely grown in Sabouraud's medium (peptone 10 g, glucose 20 g and yeast extract 5 g, per liter). To induce hyphae formation, the cells were grown in RPMI1640 medium at 37°C.Reagents 1,4-Diamino-2-butanone (DAB) was pur-The Amount of Polyamines in C. albicans C. albicans cells (1ϫ10 5 cells/ml in RPMI1640) were incubated with DAB (0, 960 mM) in 5% CO 2 at 37°C for 1 and 3 h. Cells were washed twice with sterile water and disrupted with a Mini-Beadbeater (diameter of glass beads, 0.3 mm, Central Scientific Commerce, Inc.). Total proteins in the supernatant were analyzed at 280 nm and polyamines levels were determined using a Labosearch polyamine auto (A and T Corp.).Yeast Induction of C. albicans by Polyamine Synthesis Inhibitor C. albicans cells were adjusted to 1ϫ10 5 cells/ml in RPMI1640. DAB (0, 30, 60, 120, 240, 480, 960 mM) was added and incubated in 5% CO 2 at 37°C for 3 h. The ratio of hyphae cells was counted microscopically.Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Gene arrangement participated in the hyphae formation of C. albicans referenced using the Entrez System of the National Center for Biotechnology Information (U.S.A.). PCR primers were designed using the Search Launcher in the Human Genome Sequencing Center (U.S.A.). C. albicans cells (1ϫ10 5 cells/ml in RPMI1640) were incubated with DAB (0, 960 mM) in 5% CO 2 at 37°C for 3 h. The cells were washed twice with sterile water. The cDNA library of the cells was synthesized with a Cells-tocDNA kit (Ambion Inc., U.S.A.). PCR reaction was conducted with a KOD-plus kit (Toyobo Co., Ltd.). Primers used in this assay are shown in Table 1. PCR products were analyzed using an Agilent 2100 Bioanalyzer (Agilent Technologies). The result was shown as a relative expression of mRNA compared with that of ACT1 (actin) as described by McCreach's method. 11)cAMP Concentration in C. albicans C. albicans cells (1ϫ10 5 cells/ml in RPMI1640) were incubated with DAB (0, 960 mM) in 5% CO 2 at 37°C for 3 h. Cells were washed twice with sterile water and disrupted with a Mini-Beadbeater. Total proteins in the s...

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Structure and regulation of theCandida albicans ADH1 gene encoding an immunogenic alcohol dehydrogenase

Bertram

Swoboda

Gooday

et al. 1996

Yeast

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The Candida albicans ADH1 gene encodes an alcohol dehydrogenase which is immunogenic during infections in humans. The ADH1 gene was isolated and sequenced, and the 5′‐ and 3′‐ends of its mRNA were mapped. The gene encodes a 350 amino acid polypeptide with strong homology (70·5–85·2% identity) to alcohol dehydrogenases from Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. The cloned C. albicans ADH1 gene was shown to be functional through complementation of adh mutations and efficient production of active alcohol dehydrogenase in S. cerevisiae. Northern analysis of C. albicans RNA revealed that ADH1 mRNA levels were regulated in response to carbon source and during batch growth. During growth on glucose, ADH1 mRNA levels rose to maximum levels during late exponential growth phase and declined to low levels in stationary phase. The ADH1 mRNA was relatively abundant during growth on galactose, glycerol, pyruvate, lactate or succinate, and less abundant during growth on glucose or ethanol. Alcohol dehydrogenase levels did not correlate closely with ADH1 mRNA levels under the growth conditions studied, suggesting either that this locus is controlled at both transcriptional and post‐transcriptional levels, or that other differentially regulated ADH loci exist in C. albicans.

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Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans

Cited by 24 publications

References 31 publications

Regulation of chitin synthesis during dimorphic growth of Candida albicans

Regulation of chitin synthesis during dimorphic growth of Candida albicans

Hyphae Formation of Candida albicans Is Regulated by Polyamines

Structure and regulation of theCandida albicans ADH1 gene encoding an immunogenic alcohol dehydrogenase

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