Analysis of Unc-51 protein kinase and related factors involved in axonal elongation
| Project/Area Number |
07458185
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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 |
Molecular biology
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
OHSHIMA Yasumi Kyushu Univ., Dept.of Biology, Professor, 理学部, 教授 (90037606)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1996: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1995: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | C.elegans / neuronal circuit / axonal elongation / unc-51 gene / unc-14 gene / protein kinase / axonal guidance / 線虫C.elegans / 神経軸索 / 酵母two-hybrid系 / HSP90 / EB1 |
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| Research Abstract |
Certain unc mutants in the nematode Caenorhabditis elegans, such as unc-14 and unc-51, show abnormal axonal elongation and axonal structures. Previously, we cloned the unc-51 gene and predicted that it encodes a novel serine/threonine protein kinase. In this study, we precisely localized the activity to rescue an unc-14 mutation. Also, we identified four cDNA clones encoded by the unc-14 rescuing region, in screens for proteins that bind to UNC-51 using a yeast two hybrid system. A mutation site in the cDNA was identified for each of the six unc-14 mutants, establishing that the nuc-14 gene was cloned. The unc-14 gene encodes a novel protein of 665 amino acids, and is co-expressed with the unc-51 gene in the cell bodies and axons of almost all neurons including DD/VD and HSN neurons. Another clone recovered in the two hybrid screen encodes a C-terminal region of UNC-51. Analysis using the yeast two hybrid system suggested that a central region of UNC-14 (amino acid residues 200-382) bound to a C-terminal region of UNC-51 (455-856), and that the UNC-51 C-terminal region oligomerized. In in vitro binding studies using recombinant fusion proteins produced in E.coli, UNC-14 directly interacted with UNC-51. We propose that UNC-51 protein kinase acts as an oligomer, and that UNC-14 is a regulator of UNC-51, in the axonal elongation and guidance.
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Report
(3 results)
Research Products
(10 results)
