Functional Analysis of an Anchoring Protein CG-NAP in the Golgi Apparatus.
| Project/Area Number |
15570159
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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 |
Cell biology
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| Research Institution | Kobe University |
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Principal Investigator |
TAKAHASHI Mikiko Kobe University, Biosignal Research center, Assistant professor, バイオシグナル研究センター, 助手 (90324938)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | Golgi apparatus / membrane traffic / coiled-coil protein / microtubule / centrosome / phosphorylation |
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| Research Abstract |
CG-NAP is a giant coiled-coil protein localized at centrosome and the Golgi apparatus. To investigate the role of CG-NAP in the Golgi apparatus, the Golgi-targeting mechanism was analyzed and the effects of dissociation of CG-NAP from the Golgi were examined on the Golgi morphology and on the membrane traffic.
1.By immunofluorescence microscopy of CG-NAP in intact and semi-intact cells, it was revealed that CG-NAP begins to dissociate from the Golgi at late G2 phase and completely dissociates at prophase.
2.The Golgi-targeting region was identified to the amino-terminal region of CG-NAP by examining the localization of various deletion mutants. This region was effectively phosphorylated by mitotically activated protein kinase plk, which might cause dissociation of CG-NAP from the Golgi.
3.Overexpression of the Golgi-targeting region resulted in dissociation of endogenous CG-NAP from the Golgi in COS cells. In these cells, Golgi marker proteins represented fragmented localization, suggesting that the Golgi apparatus is fragmented. Further, GFP-tagged Golgi marker proteins showed unstable movements in live-cell imaging. On the other hand, the transport of VSV-Gts protein from endoplasmic reticulum to plasma membrane was not inhibited.
4.When CG-NAP expression was suppressed by siRNA, similar effects were observed on the Golgi morphology and on the VSV-Gts traffic.
These results suggest that CG-NAP is involved in the formation and/or maintenance of the Golgi apparatus at pericentrosomal area.
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Report
(3 results)
Research Products
(11 results)
