| Project/Area Number |
16370005
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Genetics/Genome dynamics
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| Research Institution | OKAYAMA UNIVERSITY |
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Principal Investigator |
KAGAWA Hiroaki Okayama University, Graduate School of Natural Science and Technology, Professor, 大学院自然科学研究科, 教授 (10022732)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥12,700,000 (Direct Cost: ¥12,700,000)
Fiscal Year 2006: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2005: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2004: ¥6,400,000 (Direct Cost: ¥6,400,000)
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| Keywords | Caenorhabditis elegans / body wall muscle gene / transcription factor / troponin C / green fluorescence protein / yeast one hybrid system / MyoD / xyxsin / GFP(蛍光蛋白質) / RNAi(RNA干渉法) / ウェスタン解析 / 筋肉潰伝子 |
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| Research Abstract |
MyoD homolog of Caenorhabditis elegans is important but not essential for the body-wall muscle gene expression. Three enhancer sequences of the CeMyoD gene, hlh-1 are isolated in the second intron of the gene. To isolate the regulatory factor of body-wall muscle gene expression, which determines quantity of muscle proteins and time of muscle formation, we performed a modified-yeast one-hybrid screening and isolated LIM-8 and CeTIS-11 that show homology to TRIP6/Zyxin and TIS-11, respectively. LIM-8 interacted with CeMyoD but did not bind to enhancer sequences directly. A series of lim-8::gfp fusion genes were expressed in body-wall, vulval and anal muscles, and other non-muscle tissues. Anti-LIM-8 stained the locus of non-overlapping to DAPI staining DNA region in nucleus of body wall muscles. CeTIS-11 was also located body wall muscles and other tissues. lim-8 (RNAi) or CeTIS-11 (RNAi) worm decreased to 80% or 18% of the mobility of non-treated animal. CeTIS-11 (RNAi) worm also show egg-laying defect that comes from loss function of vulva muscle. This was the difference between LIM-8 bound to CeMyoD and CeTIS-11 directly bound to 1275 enhancer of the CeMyoD gene. These results suggest that LIM-8 regulates the body-wall muscle gene expressions interacting with CeMyoD. We propose an expression control model for the body wall genes.
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