Molecular Basis of Fruiting-body Formation in Basidiomycete Fungus

担子菌子实体形成的分子基础

• 批准号: 13660084
• 负责人: SHISHIDO Kazuo
• 金额: $ 2.05万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660084/
• 关键词: basidiomycete; fruiting-body formation; DNA-binding activity; transcription factor; target gene; gill tissue (spore-forming organ); 昆虫細胞; タンパク質間相互作用; 細胞接着

Fruiting-body formation of basidiomycetes is one of the most typical and attractive morphological differentiation observed in microorganisms. To understand the molecular mechanisms of fruiting-body formation in Lentinula edodes (Shiitake), we have studied on the target genes of two DNA-binding transcription factors PRIB and Le.CDC5, both of which regulate the fruiting-body formation. The PRIB protein binds to a 16-bp sequence with the consensus sequence of 5'GGGGGGGACAGGANCC3' and the Le.CDC5 protein binds to a 7-bp sequence with theconsensus sequence 5'GCAATGT3'. MfbA is also interesting protein that regulates the fruiting-body formation and possesses RGD sequence, PHD finger and SET domain. So we studied properties of MfbA protein. Concerning the PRIB, Le.CDC5 and MfbA, we obtained the following results. (1)The three target genes were identified for PRIB protein, ie., priB, uck1 (UMP-CMP kinase genes) and mfbC, the homologue of Saccharomyces cerevisiae YJRO070c. The mfbC gene is cons … More idered to encode the protein interacting to a putative translation initiation factor eIF5A. Only mature fruiting body was shown to contain the transcript of the mfbC gene almost exclusively, suggesting that mfbC may play a role at the final stage of fruiting-body formation. The mfbC transcript showed a region-specific distribution in hymenophore (gill tissue). High level of the mfbC transcript was detected in the outer region of trama, which is the region branching out into subhymenium. (2)We isolated two (3.7-kb and 3.9-kb) genomic HindIII fragments, to which Le.CDC5 protein binds. The two fragments contained several 7-bp consensus-like sequences and Le.CDC5 protein bound to them. They contained two or three genes of which functions were unknown. (3)The PHD finger-containing fragment of MFBA protein was found to bind to a 7-bp sequence with the consensus sequence of 5'ANAAGGT3'. Computer searching gave the information that the SET domain may be related to the methylase activity for the lysine residues of histone H3. These imply the possibility that MFBA protein may be a sort of chromatin-remodelling protein. Less

担子菌的子实体形成是微生物中最典型和最吸引人的形态分化之一。为了了解香菇子实体形成的分子机制，我们对两个dna结合转录因子PRIB和Le的靶基因进行了研究。CDC5，两者都调节子实体的形成。PRIB蛋白与一个16 bp的序列结合，其一致序列为5‘GGGGGGGACAGGANCC3’和Le。CDC5蛋白与一个7 bp的序列结合，其一致序列为5‘GCAATGT3’。MfbA是调控子实体形成的重要蛋白，具有RGD序列、PHD指和SET结构域。因此，我们研究了MfbA蛋白的性质。关于PRIB， Le。CDC5和MfbA，我们得到了以下结果。(1)鉴定出PRIB蛋白的3个靶基因，即；， priB, uck1 （UMP-CMP激酶基因）和mfbC，酿酒酵母YJRO070c的同源物。mfbC基因被认为可以编码与翻译起始因子eIF5A相互作用的蛋白。只有成熟的子实体几乎完全含有mfbC基因的转录本，这表明mfbC可能在子实体形成的最后阶段发挥作用。mfbC转录本在膜膜（鳃组织）中呈现区域特异性分布。在创伤的外部区域检测到高水平的mfbC转录本，这是分支到膜下层的区域。(2)我们分离到了两个（3.7 kb和3.9 kb）的基因组HindIII片段。CDC5蛋白结合。这两个片段含有多个7bp的consensus-like序列和Le。CDC5蛋白与它们结合。它们含有两三个功能未知的基因。(3)发现MFBA蛋白PHD指状片段与7 bp序列结合，一致序列为5’anaaggt3’。计算机检索显示SET结构域可能与组蛋白H3赖氨酸残基的甲基化酶活性有关。这暗示MFBA蛋白可能是一种染色质重塑蛋白。少

项目成果

Kazuo Shishido: "Two DNA-binding transcription factors involved in fruiting-body formation in the basidiomycete Lentinula edodes and their target genes."Abstract of XXII Fungal Genetics Conference (March 18-23, Asilomar). 78 (2003)

Kazuo Shishido：“两种 DNA 结合转录因子参与担子菌香菇子实体的形成及其靶基因。”第二十二届真菌遗传学会议摘要（3 月 18 日至 23 日，Asilomar）。

宍戸 和夫, 他: "担子菌きのこの子実体形成の分子機構(シンポジウム"生物の形つくり")"日本農芸化学会2002年度大会(2002年3月)講演要旨集. 431 (2002)

Kazuo Shishido 等人：“担子菌子实体形成的分子机制（研讨会“生命的塑造”）”日本农业化学学会 2002 年年会摘要集（2002 年 3 月 431）。 ）

Y.Miyazaki, Y.Sakuragi, T.Yamazaki, K.Shishido: "Target genes of the developmental regulator PRIB of the mushroom Lentinula edodes"Biosci.Biotechnol.Biochem.. (in press). (2004)

Y.Miyazaki、Y.Sakuragi、T.Yamazaki、K.Shishido：“香菇发育调控 PRIB 的目标基因”Biosci.Biotechnol.Biochem..（出版中）。

宍戸 和夫 他: "担子菌きのこの子実体形成の分子機構(シンポジウム"生物の形づくり")"日本農芸化学会2002年度大会講演要旨集. 431 (2002)

Kazuo Shishido 等人：“担子菌子实体形成的分子机制（研讨会“生物的塑造”）”日本农业化学学会 2002 年年会摘要 431（2002）。

Y.Miyazaki, T.Jojima, T.Ono, T.Yamazaki, K.Shishido: "A cDNA honiologue of Schizosaccharomyces pombe cdc5 from the Mushroom Lentinula edodes : characterization of the cDNA and its expressed product"Biochimica et Biophysica Acta. (submitted).

Y.Miyazaki、T.Jojima、T.Ono、T.Yamazaki、K.Shishido：“来自蘑菇香菇的粟酒裂殖酵母 cdc5 的 cDNA 同源物：cDNA 及其表达产物的表征”Biochimica et Biophysicala Acta。