De Groot P W, Schaap P J, Sonnenberg A S, Visser J, Van Griensven L J
Wageningen Agricultural University, The Netherlands.
J Mol Biol. 1996 Apr 19;257(5):1008-18. doi: 10.1006/jmbi.1996.0219.
Differential screening of a cDNA library was used to clone genes that are specifically expressed during mushroom development in the basidiomycete Agaricus bisporus. One of the isolated genes encodes a polypeptide of 112 amino acid residues and belongs to the fungal gene family encoding hydrophobins. This gene, hypA, has the characteristic pattern of eight cysteine residues at conserved positions and a hydrophobicity pattern that is very similar to class I hydrophobins. Elucidation of the genomic structure of hypA led to the identification of a second copy, hypC, located downstream of hypA. Although at a much lower level, hypC is like hypA specifically expressed on fruit bodies. The hypA mRNA level is transiently increased ten days after fruit body induction and expression appears to be associated with rapid expansion of the mushroom caps. In mushroom caps, very high concentrations of hypA messengers were found in the (outer) peel tissue, where they accumulate to more than 60% of the total mRNA mass. The corresponding protein with a molecular mass of 8 to 9 kDa was purified from this peel tissue and was identified by N-terminal sequencing. Our results suggest that HYPA forms a protective hydrophobic layer instrumental in cap formation.
利用对cDNA文库的差异筛选来克隆在担子菌双孢蘑菇子实体发育过程中特异性表达的基因。其中一个分离出的基因编码一个由112个氨基酸残基组成的多肽,属于编码疏水蛋白的真菌基因家族。这个基因,hypA,在保守位置具有八个半胱氨酸残基的特征模式以及与I类疏水蛋白非常相似的疏水性模式。对hypA基因组结构的阐明导致鉴定出位于hypA下游的第二个拷贝hypC。尽管水平低得多,但hypC与hypA一样在子实体上特异性表达。子实体诱导十天后,hypA mRNA水平短暂升高,且表达似乎与蘑菇菌盖的快速扩展有关。在蘑菇菌盖中,在(外层)果皮组织中发现了非常高浓度的hypA信使RNA,它们在那里积累到总mRNA量的60%以上。从该果皮组织中纯化出分子量为8至9 kDa的相应蛋白质,并通过N端测序进行了鉴定。我们的结果表明,HYPA形成了一个有助于菌盖形成的保护性疏水层。