Physiological Analysis of Mutants Indicates Involvement of the

This paper reports a phenotypic characterization of ggpl mutants. The cloned GGPI (GAS)) gene, which encodes a mawjor GPI-anchored glycoprotein (gpllS) of Saccharomyces cerevisiae of unknown function, was used to direct the inactivation of the chromosomal gene in haploid and diploid strains by gene replacement. The analysis of the null mutants reveals a reduction in the growth rate of 15 to 40%. Cells are round, with more than one bud, and extensively vacuolized. In the stationary phase, mutant cells are very large, arrest with a high percentage of budded cells (about 54 and 70%o for haploid and diploid null mutants, respectively, in comparison with about 10 to 13 % for control cells), and have reduced viability. The observed phenotype suggests defects in cell separation. Flow cytometric analysis of DNA reveals an increase in the fraction of cells in the G2+M+G1 * compartment during exponential growth. Conjugation and sporulation are not affected. The exocellular location of gpll5 led us to examine cell wall properties. Cell wall and septum ultrastructure of abnormally budded cells was analyzed by electron microscopy analysis, and no appreciable differences from wild-type cells were found. Microscopic analysis revealed an increase in chitin content and delocalization. In comparison with control cells, ggpl null mutants are shown to be resistant to Zymolyase during the exponential growth phase. A fivefold overexpression of gpll5 does not bring about any effects on cell growth parameters and cell wall properties