1887

Abstract

Summary: A secreted phosphate-repressible acid phosphatase from has been purified and the N-terminal region and an internal peptide have been sequenced. Using synthetic oligodeoxyribonucleotides based on the sequenced regions, the genomic sequence, , encoding the protein has been isolated. The deduced protein, named KIPho5p, consists of 469 amino acids and has a molecular mass of 52 520 Da (in agreement with the data obtained after treatment of the protein with endoglycosidase H). The purified enzyme shows size heterogeneity, with an apparent molecular mass in the range 90-200 kDa due to the carbohydrate content (10 putative glycosylation sites were identified in the sequence). A 16 amino acid sequence at the N-terminus is similar to previously identified signal peptides in other fungal secretory proteins. The putative signal peptide is removed during secretion since it is absent in the mature secreted acid phosphatase. The gene can be induced 400-600-fold by phosphate starvation. Consensus signals corresponding to those described for - and -binding sites are found in the 5′ region. Northern blot analysis of total cellular RNA indicates that the gene codes for a 1.8 kb transcript and that its expression is regulated at the transcriptional level. Chromosomal hybridization indicated that the gene is located on chromosome II. The gene of is able to functionally complement a mutation of Southern blot experiments, using the gene as probe, show that some reference strains lack repressible acid phosphatase, revealing a different gene organization for this kind of multigene family of proteins as compared to

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1997-08-01
2024-10-03
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