2003 Fiscal Year Final Research Report Summary
Functional analysis of Sec1/Munc-18 family involved in membrane traffic
| Project/Area Number |
14580748
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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 |
Neurochemistry/Neuropharmacology
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| Research Institution | Tokyo Women's Medical University School of Medicine |
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Principal Investigator |
ANDO Keiko Tokyo Women's Medical University, School of Medicine, assistant, 医学部, 助手 (40221741)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Sec1 / Munc-18 / Caenorhabditis elegans / vps45 / lysosome / SNARE |
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| Research Abstract |
The docking/fusion of transport vesicles mediated by the SNARE proteins is thought to be regulated by Sec1/Munc-18 (SM) proteins. The function of SM proteins is less well understood but their importance is emphasized by the blocks of general membrane trafficking and neurotransmitter release caused by mutations in SM proteins. There are 5 members of SM genes on the C.elegans genome, in addition to unc-18 regulating neurotransmitter release. To investigate systematically molecular structure and function of the Sec1/Munc-18 in multicellular organism, we isolated deletion mutations of all C.elegans SM proteins by TMP/UV method (K.Gengyo-Ando and S.Mitani, 2000). We are focusing on the C.elegans vps45 (Ce-vps45) whose orthologous protein in yeast plays an important role in membrane trafficking from the Golgi to vacuole.
In this work, we reported that the putative null mutation of Ce-vps45, tm246, showed a maternal-effect as larval lethal phenotype, and this phenotype was rescued when Ce-vps45 cDNA was specifically expressed in the intestine. To visualize specific membrane trafficking pathway, we performed transgenic analyses using several fluorescent reporters in the mutant background. We found that tm246 mutation blocked the targeting of cathepsin D/EGFP fusion protein to gut granules. These results suggested that Ce-vps45 gene products may function in membrane trafficking from the Golgi to lysosome in mainly intestinal cells where lysosome is highly developed. Ce-vps45 may also be required for endocytic pathway because tm246 mutant showed a coelomocyte uptake defective (Cup) phenotype (H.Fares and I.Greenwald, 2001) in restrictive temperature. Our data demonstrate that Ce-vps45 regulating two different membrane trafficking events in C.elegans.
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Research Products
(13 results)
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[Publications] Wang X, Wu YC, Fadok VA, Lee MC, Gengyo-Ando K, Cheng LC, Ledwich D, Hsu PK, Chen JY, Chou BK, Henson P, Mitani S, Xue D.: "Cell corpse engulfment mediated by C.elegans phosphatidylserine receptor through CED-5 and CED-12."Science. 302(5650). 1563-1566 (2003)
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[Publications] Kimura Y, Corcoran EE, Eto K, Gengyo-Ando K, Muramatsu MA, Kobayashi R, Freedman JH, Mitani S, Hagiwara M, Means AR, Tokumitsu H.: "A CaMK cascade activates CRE-mediated transcription in neurons of Caenorhabditis elegans."EMBO Re. 3(10)(Epub 2002 Sep 13). 962-966 (2002)
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