Molecular mechanisms of the initial stages of sexual development in the basidiomycete Coprinus cinereus
Grant-in-Aid for Scientific Research (C)
|Allocation Type||Single-year Grants|
|Research Institution||OKAYAMA UNIVERSITY|
KAMADA Takashi Okayama University, Faculty of Science, Professor, 理学部, 教授 (40033360)
|Project Period (FY)
2001 – 2003
Completed(Fiscal Year 2003)
|Budget Amount *help
¥3,900,000 (Direct Cost : ¥3,900,000)
Fiscal Year 2003 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 2002 : ¥900,000 (Direct Cost : ¥900,000)
Fiscal Year 2001 : ¥2,000,000 (Direct Cost : ¥2,000,000)
|Keywords||Basidiomycetes / Coprinus cinereus / Sexual development / Matins-tune genes / Dikaryon / Nuclear migration / きのこ / クランプ結合 / 高配型遺伝子 / ニ核菌糸|
Molecular mechanisms of the initial stages of sexual development in the basidiomycete Coprinus cinereus were studied with the following results.
1. We found that progeny isolated from a fruiting body collected in the field exhibit a distinctive mycelial development in common A matings. Genetic analysis suggested that the common A heterokaryotic phenotype is brought about by a nuclear factor(s) other than the mating type genes.
2. We identified the whole structure of the num1 gene, which is involved in nuclear migration for dikaryosis, a major B-regulated developmental process.
3. We screened a cDNA library for clones that encode proteins interacting with Num1, which identified the nip1 gene.
4. We analyzed the promoter region of the clp1 gene and identified a 10-bp sequence (5'-GATGCAAACA-3') located 155-146 upstream of the transcriptional start site, which is involved in the transcriptional control of clp1 In the future, we plan to examine whether the heterodimer of the A mating-type gene products (HD1 and HD2), which activates clp1 expression, binds the 10-bp sequence.
5. We found that Clp1 and Pcc1, both of which are working in the A-regulated developmental pathway, are interacting with each other, as examined by yeast two-hybrid assays. Furthermore, we identified that the C-terminal region of Clp1 and the N-terminal region of Pcc1 are involved in the interaction in yeast.
6. To elucidate downstream events of pcc1, we performed a subtraction experiment using the pcc1-4 mutant strain (5337#4), in which the A-regulated developmental pathway is constitutively activated, and its parental wild-type homokaryon (5337), and selected 64 cDNAs from genes possibly up-regulated in strain 5337#4. In the future, we plan to confirm differential transcription between strains 5335 and 5337#4 for each cDNA by northern analysis and then analyze the structure and function of genes expressed differentially between 5337 and 5337#4.
Research Products (14results)