Related Experiment Video

Updated: Aug 12, 2026

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
13:08

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass

Published on: August 10, 2017

Reconstitution of the Golgi reassembly process in semi-intact MDCK cells

F Kano1, K Nagayama, M Murata

  • 1Department of Biophysics, Graduate School of Science, Kyoto University, Japan.

Biophysical Chemistry
|June 14, 2000
PubMed

Insights

Golgi apparatus membranes fragment during mitosis. Ilimaquinone (IQ) causes Golgi vesiculation, and its removal triggers reassembly, revealing key factors like cytoplasmic dynein in Golgi membrane dynamics.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The Golgi apparatus disassembles into dispersed fragments during mitosis.
  • Ilimaquinone (IQ), a sea sponge metabolite, induces Golgi membrane vesiculation.
  • IQ removal allows vesiculated Golgi membranes to reassemble into stacks.

Purpose of the Study:

  • To investigate the mechanism of Golgi membrane dynamics during mitosis.
  • To reconstitute and analyze Golgi reassembly from IQ-induced vesiculated membranes.
  • To identify the biochemical factors involved in Golgi reformation.

Main Methods:

  • Utilized streptolysin O-permeabilized MDCK cells for Golgi reassembly experiments.
  • Employed a green fluorescence protein (GFP)-tagged protein to monitor Golgi membrane dynamics.
  • Performed morphometric analysis to study reassembly kinetics and biochemical requirements.

Main Results:

  • Dissected Golgi reassembly into two stages: vesiculation to punctate structures and reformation into stacks.
  • Identified an NEM-sensitive factor, cytoplasmic dynein, and a GTP-binding protein as crucial for Golgi reassembly.
  • Quantified the kinetics and biochemical dependencies of the reassembly process.

Conclusions:

  • The study elucidates the two-step mechanism of Golgi apparatus reassembly post-mitosis.
  • Key molecular players, including cytoplasmic dynein and an NEM-sensitive factor, are essential for Golgi reformation.
  • This reconstituted system provides insights into Golgi membrane dynamics and organelle biogenesis.

More Related Videos

In Vitro Reconstitution of the Actin Cytoskeleton Inside Giant Unilamellar Vesicles10:19

In Vitro Reconstitution of the Actin Cytoskeleton Inside Giant Unilamellar Vesicles

Published on: August 25, 2022

In vitro Reconstitution of Cytoskeletal Networks inside Phase Separated Giant Unilamellar Vesicles (GUVs)06:34

In vitro Reconstitution of Cytoskeletal Networks inside Phase Separated Giant Unilamellar Vesicles (GUVs)

Published on: June 20, 2025

Related Concept Videos

Golgi Apparatus01:49

Golgi Apparatus

As they leave the Endoplasmic Reticulum (ER), properly folded and assembled proteins are selectively packaged into vesicles. These vesicles are transported by microtubule-based motor proteins and fuse together to form vesicular tubular clusters, subsequently arriving at the Golgi apparatus, a eukaryotic endomembrane organelle that often has a distinctive ribbon-like appearance.The Golgi apparatus is a major sorting and dispatch station for the products of the ER. Newly arriving vesicles enter...
Vesicular Tubular Clusters01:45

Vesicular Tubular Clusters

After budding out from the ER membrane, some COPII vesicles lose their coat and fuse with one another to form larger vesicles and interconnected tubules called vesicular tubular clusters or VTCs. These clusters constitute a compartment at the ER-Golgi interface known as ERGIC (Endoplasmic Reticulum Golgi Intermediate Compartment). The ERGIC is a mobile membrane-bound cargo transport system that sorts proteins secreted from ER and delivers them to the Golgi.
With the help of motor proteins such...
Golgi Apparatus01:09

Golgi Apparatus

Properly folded and assembled proteins are selectively packaged into vesicles that exit the ER. Motor proteins transport these vesicles to the Golgi apparatus for adding modifications that make these proteins functional at their destination.
The Golgi apparatus is a eukaryotic organelle that has a distinctive ribbon-like appearance. It is a primary sorting and dispatch station for cargo arriving from the ER. Newly arriving vesicles enter the cis face of the Golgi, closest to the ER, and are...
Golgi Matrix Proteins01:12

Golgi Matrix Proteins

Golgi matrix proteins are a group of highly dynamic proteins that maintain the stacked structure of Golgi. These proteins adapt to rapid morphological changes of the Golgi during the cell cycle. During cell division, mild proteolysis removes these connections resulting in Golgi unstacking. In The daughter cells, these proteins help reassemble the unstacked Golgi.
One of the first identified Golgi matrix proteins was GM130, a rod-like protein located in the cis-Golgi. Subsequently, many Golgi...
Transport Across the Golgi01:26

Transport Across the Golgi

While it is unclear how molecules move between adjacent Golgi cisternae, it is apparent that the molecules move from cis- cisterna, the entry face, to the trans- cisterna, the exit face. Experiments initially suggested vesicles that bud from one cisterna and fuse with the next cisterna to transport proteins between the cisternae. This vesicular transport model describes the Golgi apparatus as a relatively static structure with a unique enzyme composition in each cisterna. Molecules are...
Golgi Apparatus01:09

Golgi Apparatus

Properly folded and assembled proteins are selectively packaged into vesicles that exit the ER. Motor proteins transport these vesicles to the Golgi apparatus for adding modifications that make these proteins functional at their destination.
The Golgi apparatus is a eukaryotic organelle that has a distinctive ribbon-like appearance. It is a primary sorting and dispatch station for cargo arriving from the ER. Newly arriving vesicles enter the cis face of the Golgi, closest to the ER, and are...