Journal
NATURE PROTOCOLS
Volume 5, Issue 4, Pages 758-772Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2010.38
Keywords
-
Categories
Funding
- Pardee Cancer Research Foundation
- National Institute of Health [GM087364]
- American Cancer Society [RGS-09-278-01-CSM]
- University of Michigan Rackham Faculty Research
- Michigan Alzheimer's Disease Research Center [P50 AG08761]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM087364] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE ON AGING [P01AG008761] Funding Source: NIH RePORTER
Ask authors/readers for more resources
The Golgi apparatus undergoes extensive disassembly during mitosis and reassembly in post-mitotic daughter cells. This process has been mimicked in vitro by treating Golgi membranes with mitotic and interphase cytosol. To determine the minimal machinery that controls the morphological change, we have developed a defined Golgi disassembly and reassembly assay that reconstitutes this process using purified proteins instead of cytosol. Treatment of Golgi membranes with mitotic kinases and COP I coat proteins efficiently disassembles the membranes into mitotic Golgi fragments, whereas further incubation with p97 or N-ethylmaleimide-sensitive factor (two AAA ATPases involved in membrane fusion) and their cofactors, in combination with protein phosphatase PP 2A, leads to reassembly of the membranes into new Golgi stacks. The whole process takes 3-4 d and is applicable for identification and determination of novel cytosolic and membrane proteins that regulate Golgi membrane dynamics in the cell cycle.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available