There is a debate over how protein trafficking is performed through the Golgi apparatus1,2,3,4. In the secretory pathway, secretory proteins that are synthesized in the endoplasmic reticulum enter the early compartment of the Golgi apparatus called cis cisternae, undergo various modifications and processing, and then leave for the plasma membrane from the late (trans) cisternae. The cargo proteins must traverse the Golgi apparatus in the cis-to-trans direction. Two typical models propose either vesicular transport or cisternal progression and maturation for this process. The vesicular transport model predicts that Golgi cisternae are distinct stable compartments connected by vesicular traffic, whereas the cisternal maturation model predicts that cisternae are transient structures that form de novo, mature from cis to trans, and then dissipate. Technical progress in live-cell imaging has long been awaited to address this problem. Here we show, by the use of high-speed three-dimensional confocal microscopy, that yeast Golgi cisternae do change the distribution of resident membrane proteins from the cis nature to the trans over time, as proposed by the maturation model, in a very dynamic way.
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We thank all the members of the Nakano group of the Dynamic-Bio Project for the accomplishment of this microscopy project; Yokogawa Electric Corporation, NHK (Japan Broadcasting Corporation), NHK Engineering Service, Hitachi Kokusai Electric, and the Research Association for Biotechnology for their contributions; R. Tsien for the distribution of mRFP; Olympus Corporation for technical help; and B. Glick for exchange of information before publication. This work was supported by national funds from the Ministry of Economy, Trade and Industry of Japan and the New Energy and Industrial Technology Development Organization, partly by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and partly by the funds from the Bioarchitect, the Real-Time Bionanomachine and the Extreme Photonics Projects of RIKEN.
Wild-type yeast cells expressing GFP–Rer1p (cis, green) and mRFP-Gos1p (medial, red). 20x real time.
ret1-1 cells expressing GFP–Gos1p (medial,green) and Sec7p–mRFP (trans, red). 10x real time.
Another example of ret1-1 cells expressing GFP–Gos1p (medial,green) and Sec7p–mRFP (trans, red). 10x real time.
3D observation of wild-type yeast cells expressing mRFP–Sed5p (cis, red) and Sec7p–GFP (trans, green). 5x real time.
Another example of 3D movie showing wild-type yeast cells expressing mRFP-Sed5p and Sec7p–GFP. 14x real time.
3D deconvolution observation of yeast expressing mRFP–Gos1p and Sec7p–GFP. 25x real time.
Another example of 3D deconvolution movie. 25x real time.
About this article
Scientific Reports (2016)