Function of cell-adhesion protein MFBA in fruiting-body formation of basidiomycete

细胞粘附蛋白MFBA在担子菌子实体形成中的作用

• 批准号: 07660102
• 负责人: SHISHIDO Kazuo
• 金额: $ 1.41万
• 依托单位: TOKYO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660102/
• 关键词: basidiomycete; fruiting body formation; cell-adhesion protein; cell aggregation

Fruiting of basidiomycetes is one of the most typical morphogenetic phenomena observed in eukaryotic microorganisms. We so far isolated new gene mfbA which is adjacent to ras gene and expressed strongly in mature fruiting bodies. mfbA is also expressed in immature fruiting bodies. The mfbA cDNA cloned consisted of 6700 bp encoding 2157 amino acids. MFBA protein (MW234,494) contained the cell surface attachment-promoting amino acid sequence Arg-Gly-Asp (RGD) observed first in fibronectin.This research aimed to assess the function of MFBA.Although we failed in producing intact MFBA protein, a 425-amino-acid MFBA fragment containing the RGD motif (named MFBA (582-1006) peptide) was successfully producted in E.coli. By using this MFBA fragment, the following new experimental results were obtained.(1) MFBA (582-1006) peptide exhibits a cell adhesion and spreading activity toward mammalian cells via the RGD sequence.(2) The peptide causes significant in vivo aggrefation of the yeast S.cerevisiae cells via the RGD sequence.(3) The peptide causes significant in vitro aggregation of basidiomycetous hyphal cells via RGD sequence.(4) The mfbA transcript is abundant in the gill-depleted pileus and stipe, but almost no transcript is found in the gill tissue.(5) MFBA (582-1006) peptide possesses a highly hydrophobic region adjacent to the RGD motif.These results strongly suggest that the hydrophobic region of MFBA is embedded in basidiomycetous hyphal cell wall and the RGD-containing amino acid sequence interacts with the receptor protein molecules being present on the surface of other cells via the RGD sequence. These protein-protein interactions results in formation of the net structure of basidiomycetous hyphal cells. MFBA is considered to be involved in maturation of fruiting body and formation of pileus and stipe of fruiting body.

担子菌的出菇是真核微生物中最典型的形态发生现象之一。目前我们已经分离到一个新基因mfbA，它与ras基因相邻，在成熟子实体中强烈表达。mfbA也在未成熟的子实体中表达。克隆的mfbA cDNA全长6700 bp，编码2157个氨基酸。MFBA蛋白（MW 234，494）含有在纤维连接蛋白中首次发现的促进细胞表面粘附的氨基酸序列Arg-Gly-Asp（RGD），本研究旨在研究MFBA的功能，虽然我们未能成功地制备完整的MFBA蛋白，但在大肠杆菌中成功地制备了一个含有RGD基序的425个氨基酸的MFBA片段（命名为MFBA（582-1006）肽）。通过使用该MFBA片段，获得了以下新的实验结果。(1)MFBA（582-1006）肽通过RGD序列表现出对哺乳动物细胞的细胞粘附和扩散活性。(2)该肽通过RGD序列引起酵母酿酒酵母细胞的显著体内聚集。(3)该肽通过RGD序列引起担子菌菌丝细胞的显著体外聚集。(4)mfbA转录本在鳃耗竭的菌盖和柄中丰富，但在鳃组织中几乎没有发现转录本。(5)MFBA（582-1006）肽在邻近RGD基序处具有高度疏水区，这些结果有力地表明MFBA的疏水区包埋在担子菌菌丝细胞壁中，含RGD的氨基酸序列通过RGD序列与存在于其它细胞表面的受体蛋白分子相互作用。这些蛋白质-蛋白质相互作用的结果在形成的网状结构的担子菌菌丝细胞。MFBA被认为参与子实体的成熟以及子实体的菌盖和菌柄的形成。

项目成果

Toru Yasuda, Takayuki Ishizaki and Kazuo Shishido: "Molecular mechanism of fruiting body formation (XIV) Function of cell-adhesion protein MFBA." The 1996 Annual Conference of Japan Society for Bioscience, Biotechnology, and Agrochemistry. March 30-Apri 1

Toru Yasuda、Takayuki Ishizaki 和 Kazuo Shishido：“子实体形成的分子机制 (XIV) 细胞粘附蛋白 MFBA 的功能。”

Kazuo Shishido: "A fruiting body-specific cDNA,mfbAc,from the mushroom Lentinus edodes encodes a high-molecular weight cell-adhesion protein containing an Arg-Gly-Asp motif." Proceedings of Genetics and Cellular Biology of Basidiomycetes III. 13 (1995)

Kazuo Shishido：“来自蘑菇香菇的子实体特异性 cDNA，mfbAc，编码含有 Arg-Gly-Asp 基序的高分子量细胞粘附蛋白。”

Kazuo Shishido, Yasumasa Miyazaki, Toru Yasuda, Osamu Ysunoka, Osamu Kondoh, Takayuki Ishizaki and Hitoshi Amano: "Two fruiting-specific cDNAs, priBc and mfbAc, encoding a DNA-binding protein or a cell-adhesion protein, derived from Lentinus edodes. New I

Kazuo Shishido、Yasumasa Miyazaki、Toru Yasuda、Osamu Ysunoka、Osamu Kondoh、Takayuki Ishizaki 和 Hitoshi Amano：“两种果实特异性 cDNA，priBc 和 mfbAc，编码 DNA 结合蛋白或细胞粘附蛋白，源自香菇。

O.Kondoh et al.: "A fruiting body-specific cDNA,mfbAc,from the mushroom Lentinus edodes encodes a high-molecular weigth cell-adhesion protein containing an Arg-Gly-Asp motif." Gene. 154. 31-37 (1995)

O.Kondoh 等人：“来自蘑菇香菇的子实体特异性 cDNA，mfbAc，编码含有 Arg-Gly-Asp 基序的高分子量细胞粘附蛋白。”

安田 徹,石崎孝之,宍戸和夫: "真正担子菌シイタケの子実体形成機構(XIV)細胞接着蛋白質MFBAの出芽酵母に対する作用" 日本農芸化学会誌. 70(臨時増刊号). 264 (1996)

Toru Yasuda、Takayuki Ishizaki、Kazuo Shishido：“真正的担子菌香菇（XIV）的子实体形成机制以及细胞粘附蛋白 MFBA 对酿酒酵母的影响”日本农业化学学会杂志 70（特刊）。 264（1996）