The gene was identified by homology to the orthologue and encodes a predicted 1655 amino acid protein. In most cell-wall chitin is associated with primary septum formation and Bni4p is involved in tethering the Chs3p chitin synthase enzyme to the mother-bud neck by forming a bridge between a regulatory protein Chs4p and the septin Cdc10p. Bni4p shows 20 % overall identity to the Bni4p, with 73 % identity over the C-terminal 63 amino acids, which includes a putative protein phosphatase type 1 (PP1) binding domain. Northern blot analysis revealed a transcript of the expected size that was expressed in both yeast and hyphal growth forms. has more chitin in its cell wall than , and again most chitin is synthesized by Chs3p. The function of was investigated by performing a targeted gene disruption using the ‘Ura-blaster’ method to delete amino acids 1120–1611 that are essential for function. The resulting Δ/Δ null mutants formed lemon-shaped yeast cells and had a 30 % reduction in cell-wall chitin, reduced hyphal formation on solid serum-containing medium and increased sensitivity to SDS and increased resistance to Calcofluor White. The Δ/Δ null mutants were unaffected in chitin ring formation, but often exhibited displaced bud sites with more obvious but flattened birth scars. Therefore, unlike in , the mutant apparently alters chitin distribution throughout the cell wall and not exclusively at the bud-neck region.


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