| Project/Area Number |
17570179
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | Hiroshima University |
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Principal Investigator |
YAMAMOTO Takashi Hiroshima University, Graduate School of Science, Professor (90244102)
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| Co-Investigator(Kenkyū-buntansha) |
SHIBATA Tatsuo Hiroshima University, Graduate School of Science, Associate Professor (10359888)
SAKAMOTO Naoaki Hiroshima University, Graduate School of Science, Associate Professor (00332338)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,740,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥240,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | gene / development / regulation of expression / genome / 発生・分化 / 転写調節因子 |
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| Research Abstract |
The purpose of this study is to analyze the regulatory mechanisms of spatio-temporal specific gene expression during development. We focus on the T-brain homolog (HpTb) of sea urchin, Hemicentrotus pulcherrimus, which expresses specifically in micromere descendants and is involved in the gastrulation and skeletogenesis. The following results were obtained from series of experiments.
(1) We isolated approximately 7 kb genomic region of HpTb gene and showed that GFP expression driven by this region exhibits the spatio-temporal pattern corresponding substantially to that of endogenous HpTb expression.
(2)T o address the cis-regulatory region within the 7 kb region of HpTb gene, the nucleotide sequence of the 7 kb region was compared with the corresponding region of genes encoding the T-brain ortholog of other sea urchin species, Strongylocentrotus purpuratus and Lytechinus variegatus by Family RelationsII software. Four conserved regions (C1, C2, C3 and C4) were found besides exons. The Cl region locates in the 5' franking region and C2 to C4 regions in the 1st intron. By computational search, Hairy family and Snail family consensus sequences were found in C1 and C2 region, respectively
(3) Deletion of interspecifically conserved C2 and C4 regions resulted in an increase of ectopic expression. Mutations in Hairy family and Snail family consensus sequences also increased ectopic expression, suggesting that these elements are involved in repression of HpTb gene expression in ectoderm and endoderm.
(4) We demonstrated that C4 region functions as enhancer, and that three Ets family consensus sequences are involved in this activity but not in spatial regulation.
From these results, we concluded that the specific expression of HpTb gene is regulated by multiple cis-regulatory elements
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