| 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)
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| Research Abstract |
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.
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