Biomedical subjects

W L Chaffin

Publications and source records attributed to W L Chaffin.

44 records · Page 3Linked to original sources

PubMed1981

Morphological commitment in Candida albicans.

Stationary phase yeast cells of the dimorphic fungus albicans can reinitiate growth under appropriate conditions either as yeasts through bud formation or as hyphae through germ tube formation and elongation. Stationary phase yeast cells resuspended in fresh medium at 37 degrees C form germ tubes and those resuspended at 25 degrees C form buds. Temperature shift experiments have been used to observe when cells become committed to germ tube formation and yeast budding growth under conditions favorable to each form. The two commitment processes appear to be independent and, once initiated, occur at characteristic rates with commitment to germ tube formation preceding commitment to yeast bud formation. The rate of commitment to germ tube formation was consistent with a random process or first-order kinetics. A relationship between cell volume and commitment to yeast growth and bud emergence was consistent with observations of cell volume distribution both in stationary phase cultures and between budded and unbudded cells during resumption of growth at 25 degrees C.

Candida albicans↗

PubMed1981

Differential expression of cytoplasmic proteins during yeast bud and germ tube formation in Candida albicans.

Changes in the identity and quantity of proteins synthesized during morphogenesis may result from alterations in gene expression in the dimorphic yeast Candida albicans. Stationary phase yeast cells, upon resuming growth at 25 degrees C, form budding yeast and at 37 degrees C form germ tubes. In order to identify proteins associated with morphogenesis, we compared cytoplasmic proteins synthesized during germ tube and bud formation. Proteins synthesized during this period were labeled at four intervals with either [3H]leucine or [35S]methionine and separated by two-dimensional polyacrylamide gel electrophoresis. This study shows that, of the 230 proteins resolved on each gel, 5 were specific to the yeast morphology and 2 proteins showed reduction in net synthesis in the mycelial phase. There were, however, no mycelium-specific proteins at any labeling period. The majority of proteins were common to both morphologies and showed no major shift in number during resumption of growth. The observations reported here suggest that differential gene expression occurs during morphogenesis of C. albicans.

Candida albicans↗

PubMed1980

Nutrient-limited yeast growth in Candida albicans: effect on yeast-mycelial transition.

The yeast-mycelial transition in Candida albicans can be induced from yeast cells grown on minimal defined medium only in stationary phase. This study examined the inducibility of cultures in which growth was limited by the availability of the nutrients, glucose, NH4Cl, or galactose. The results showed that neither stationary phase nor cell cycle stage alone was a sufficient condition to support subsequent germ tube formation. In addition, final cell concentration alone was not a factor in inducibility. When a hundredfold decrease in growth was obtained by limiting any of the nutrients, a loss in inducibility was observed. However, the loss of inducibility differed with the limiting nutrient. Galactose, NH4Cl, and glucose-limited cultures showed respectively 15, 30, and 80% loss of inducibility. Thus the effect was associated with both carbon/energy and nitrogen-limited cells; however, glucose appeared to have a specific effect. These observations suggest that the metabolic state of the stationary phase yeast cell was an important factor in the subsequent ability to respond to conditions inducing germ tube formation.

Ammonium Chloride↗

PubMed1979

Nicotinamide adenine dinucleotide metabolism in Candida albicans.

The functional pathways of nicotinamide adenine dinucleotide (NAD) biosynthesis and their regulation were studied in the dimorphic fungus Candida albicans. The presence of a functional endogenous pathway of NAD biosynthesis from tryptophan was demonstrated. In addition, nicotinamide served as an efficient salvage precursor for NAD biosynthesis but nicotinate was not utilized. The pathway for nicotinamide utilization involved nicotinate and nicotinate nucleotides as intermediates, suggesting that the failure to utilize nicotinate involves a transport defect. The mechanisms that regulate NAD levels during exponential growth operated to maintain constant NAD levels when NAD biosynthesis occurred exclusively from endogenous or salvage pathways or from a combination of the two. The regulation also operated such that the salvage pathway was preferentially utilized.

Biological Transport↗

PubMed1976

Germ tube formation from zonal rotor fractions of Candida albicans.

Homogenous cell populations of increasing cell volume may have been isolated from exponential and stationary culture of Candida albicans by centrifugation on a sucrose gradient. Observations of the yeast-mycelial transition using these populations showed the following. (i) No fraction from early logarithmic phase cells was unable to undergo morphological transition. (ii) The time of initiation of germ tube production was correlated with cell size in stationary-phase cultures. (iii) The rate of appearance of germ tubes was nearly identical in all fractions measured. (iv) Addition of N-acetyl-D-glucosamine to homogeneous cell populations decreased the time of initial appearance of germ tubes but did not affect the rate of appearance after initiation.

Acetylglucosamine↗

PubMed1974

Nature of ribonucleic acid synthesis during early sporulation in Saccharomyces cerevisiae.

Phosphate uptake in sporulating cultures of Saccharomyces cerevisiae has been found to occur approximately 2 h after the transfer to sporulation medium. Early ribonucleic acid synthesis begins at approximately 4 h and continues to 8 h. Incorporation of phosphate into acid-extractable precursor pools parallels phosphate uptake. In triple-labeling experiments it was observed that the breakdown of vegetatively synthesized ribonucleic acid is not a significant source of precursors for ribonucleic acid synthesis during sporulation. The majority of the ribonucleic acid made in a 10-min period during sporulation does not migrate on gels with precursor or mature ribosomal ribonucleic acid.

Adenine↗

PubMedDate not supplied

Dynamic expression of cell wall proteins of Candida albicans revealed by probes from cDNA clones.

Five cDNA clones were selected from the positive clones detected by screening a germ tube expression library constructed in lambda gt11 with rabbit antisera raised against cell wall extracts of Candida albicans. The selected clones were amplified and used to obtain affinity purified antibodies by eluting from the expressed proteins that had been previously transferred onto nitrocellulose discs. The antibodies obtained were used as probes in immunoblots of the cell wall extracts separated by denaturing polyacrylamide electrophoresis. A single protein band was detected for each clone. Detection of products of the cloned sequences varied according to the extraction procedure and/or cell morphology. These products included bands exhibiting apparent molecular weights of 40, 58, 68 and 70 kDa present in beta-mercaptoethanol (beta ME) extracts from both yeast and germ tubes, and a 30 kDa beta ME extracted protein specific for germ tubes. The expression of these products at the cell surface was confirmed by indirect immunofluorescence. Expression of the mRNAs of the different cDNA clones varied according to growth- and morphology-related factors and showed no direct correlation between expression and presence in the cell wall. These observations suggest that complex mechanisms are involved in the regulation and expression of cell surface components of C. albicans.

Animals↗

PubMedDate not supplied

Cloning and characterization of a cDNA coding for Candida albicans polyubiquitin.

Immunoscreening of a Candida albicans cDNA library in the expression vector lambda gt11 with rabbit polyclonal antibodies against the 37 kDa cell surface laminin receptor of C albicans resulted in the isolation of a cDNA clone of 0.9 kb. Sequencing of this clone demonstrated a full length open reading frame encoding the polyubiquitin, which contains three tandem copies, head-to-tail spacerless repeats, of the 228 nucleotides coding for the 76 amino acids of the ubiquitin protein, which is identical to that of Saccharomyces cerevisiae. The third copy possesses an extra C-terminal amino acid which is distinct to that found in S. cerevisiae. Northern blot analysis revealed a single mRNA population of about 1 kb present in similar amounts in both yeast and mycelial cells. This indicates that the C. albicans polyubiquitin gene (UBI1) encodes a polyubiquitin precursor protein containing three ubiquitin repeats. Immunofluorescence and Western immunoblotting experiments with polyclonal antibodies against mammalian ubiquitin suggest the presence of ubiquitinated protein moieties in the wall of C. albicans cells.

Amino Acid Sequence↗