1887

Abstract

In order to identify novel genes affecting cell wall integrity, we have generated mutant strains of the filamentous fungus that show hypersensitivity to the chitin-binding agent Calcofluor White (CFW). Affected loci are designated loci. The phenotype of one of these alleles, , also includes shortened hyphal compartments and increased density of branching in the absence of CFW, as well as reduced staining of cell walls by the lectin FITC–Concanavalin A (ConA), which has strong binding affinity for mannosyl residues. We have identified two genes (AN8848.3 and AN9298.3, designated and , respectively) that complement all aspects of the phenotype. Both genes show strong sequence similarity to GDP-mannose transporters (GMTs) of and other yeasts. Sequencing of from the mutant strain reveals a G to C mutation at position 943, resulting in a predicted alanine to proline substitution at amino acid position 315 within a region that is highly conserved among other fungi. No mutations were observed in the mutant strain's allele of . Meiotic mapping demonstrated a recombination frequency of under 1 % between the locus and the locus (located ∼9.5 kb from AN8848.3), confirming that and are identical. A GmtA–GFP chimera exhibits a punctate distribution pattern, consistent with that shown by putative Golgi markers in . However, this distribution did not overlap with that of the putative Golgi equivalent marker CopA–monomeric red fluorescent protein (mRFP), which may indicate that the physically separated Golgi-equivalent organelles of represent physiologically distinct counterparts of the stacked cisternae of plants and animals. These findings demonstrate that and play roles in cell wall metabolism in similar to those previously reported for GMTs in yeasts.

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