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F Klebl

Publications and source records attributed to F Klebl.

21 records · Page 2Linked to original sources

Functional expression of the Chlorella hexose transporter in Schizosaccharomyces pombe.

Schizosaccharomyces pombe cells were transformed with an S. pombe expression vector containing a full-length cDNA of the Chlorella hexose transporter. The transformed cells accumulated 3-O-methylglucose up to 10-fold, whereas wild-type S. pombe and control transformants could only equilibrate this sugar analogue. In a pH-jump experiment, in which extracellular pH was lowered by 1.9 units, the accumulation ratio was increased in transformed cells but not in control cells. This result indicates that the gene product, Chlorella H+/glucose-symporter protein, and a pH gradient suffice for active sugar uptake. Km values for glucose, 6-deoxyglucose, and 3-O-methylglucose of 1.5 x 10(-5) M, 2.7 x 10(-4) M, and 1.0 x 10(-3) M, respectively, were identical in Chlorella and in S. pombe cells transformed with Chlorella cDNA and approximately 100-fold lower than those of the endogenous transport system of S. pombe.

Journal Article↗

Molecular cloning of a cell wall exo-beta-1,3-glucanase from Saccharomyces cerevisiae.

A major protein of Saccharomyces cerevisiae cell walls is a 29-kilodalton glycoprotein which shows lectinlike binding to beta-1,3-glucan and chitin. It was solubilized by heating isolated cell walls at 90 degrees C and purified to homogeneity by running two high-pressure liquid chromatography columns. With the sequence information of the N terminus and seven peptides, two oligonucleotides were synthesized and the gene was cloned. Its sequence is similar to those of two plant beta-glucanases, and the protein was shown to possess beta-1,3-exoglucanase activity with laminarin as substrate. Haploid yeast cells contained one copy of the gene (BGL2). Gene disruption did not result in a phenotype.

Amino Acid Sequence↗

A temperature-sensitive N-glycosylation mutant of S. cerevisiae that behaves like a cell-cycle mutant.

The temperature-sensitive S. cerevisiae mutant alg1-1, defective in the N-glycosylation of proteins, shows a first cycle arrest at the non-permissive temperature of 36 degrees C. The cell number increases by 50% and the absorbance approximately doubles. The budding index of 0.4 at 26 degrees C drops to 0.15 and DNA synthesis quickly comes to a halt at 36 degrees C. When the temperature is lowered again, budding and DNA synthesis start after a lag of 2-3 h; alpha-factor prevents both these processes in cells of mating type a. In addition, cells arrested at 26 degrees C in G1 with alpha-factor also do not start budding at the non-permissive temperature after removal of alpha-factor. The results support recent findings obtained with tunicamycin and suggest that at least one glycoprotein is required for G1-S phase transition in yeast.

DNA, Fungal↗