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Live imaging of yeast Golgi cisternal maturation

Nature volume 441, pages 1007–1010 (2006)Cite this article

Abstract

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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Figure 1: Dual-fluorescence observation of yeast Golgi cisternae.
Figure 2: Dual fluorescence observation with the αCOP mutant ret1-1.
Figure 3: Three-dimensional observation of yeast Golgi cisternae.
Figure 4: Three-dimensional deconvolution observation of yeast Golgi cisternae.

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Acknowledgements

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.

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Author notes

  1. Masaki Takeuchi

    Present address: Department of Molecular Structure, Institute for Molecular Science, National Institutes of Natural Sciences, Okazaki, Aichi, 444-8585, Japan

Authors and Affiliations

  1. Molecular Membrane Biology Laboratory, RIKEN Discovery Research Institute, Japan, Wako, Saitama, 351-0198

    Kumi Matsuura-Tokita, Masaki Takeuchi, Akira Ichihara & Akihiko Nakano

  2. Biocenter, Yokogawa Electric Corporation, Tokyo, Musashino, 180-8750, Japan

    Akira Ichihara & Kenta Mikuriya

  3. Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Bunkyo-ku, 113-0033, Japan

    Akihiko Nakano

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Correspondence to Akihiko Nakano.

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Supplementary information

Supplementary Figure 1

This figure shows a model of Golgi maturation. (PDF 440 kb)

Supplementary Figure 2

This figure shows the effects of traffic mutations on the localization of Golgi markers. (PDF 320 kb)

Supplementary Figure 3

This figure shows frequent change of color in cells expressing GFP–Gos1p (medial, green) and Sec7p–mRFP (trans, red). (PDF 909 kb)

Supplementary Video 1

Wild-type yeast cells expressing GFP–Rer1p (cis, green) and mRFP-Gos1p (medial, red). 20x real time. (MOV 2709 kb)

Supplementary Video 2

ret1-1 cells expressing GFP–Gos1p (medial,green) and Sec7p–mRFP (trans, red). 10x real time. (MOV 869 kb)

Supplementary Video 3

Another example of ret1-1 cells expressing GFP–Gos1p (medial,green) and Sec7p–mRFP (trans, red). 10x real time. (MOV 769 kb)

Supplementary Video 4

3D observation of wild-type yeast cells expressing mRFP–Sed5p (cis, red) and Sec7p–GFP (trans, green). 5x real time. (MOV 984 kb)

Supplementary Video 5

Another example of 3D movie showing wild-type yeast cells expressing mRFP-Sed5p and Sec7p–GFP. 14x real time. (MOV 621 kb)

Supplementary Video 6

3D deconvolution observation of yeast expressing mRFP–Gos1p and Sec7p–GFP. 25x real time. (MOV 926 kb)

Supplementary Video 7

Another example of 3D deconvolution movie. 25x real time. (MOV 1098 kb)

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Matsuura-Tokita, K., Takeuchi, M., Ichihara, A. et al. Live imaging of yeast Golgi cisternal maturation. Nature 441, 1007–1010 (2006). https://doi.org/10.1038/nature04737

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