In a search for genes that are only expressed in fruit bodies of the basidiomycete two cDNAs, and that encode hydrophobins have been isolated previously. In this study, the structure of is resolved and it is shown that the two genes are differentially expressed, indicating that the encoded hydrophobins serve different functions in mushrooms. encodes a polypeptide (HYPB) of 119 aa that shows little sequence identity with HYPA apart from the characteristic arrangement of eight cysteines found exclusively in hydrophobins. The temporal and spatial expression of the two hydrophobin-encoding genes during fruit body development was compared using Northern analysis and hybridization. Accumulation of mRNA was found in tissue fractions consisting of undifferentiated white hyphae. hybridization showed that the highest mRNA levels are not found in the outermost cell layers of the pileipellis but in the cell layers adjacent to that. The highest level of expression of occurs early in development when the primordium differentiates into densely packed, randomly oriented cap hyphae and loosely packed, vertically oriented stipe hyphae. In mature mushrooms, a strong accumulation of transcripts was found only in the transitional zone between cap and stipe tissue, demonstrating that transcription regulation of is clearly distinct from


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