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The GM130 and GRASP65 Golgi proteins cycle through and define a subdomain of the intermediate compartment

Nature Cell Biology volume 3, pages 11011113 (2001) | Download Citation

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Abstract

Integrating the pleomorphic membranes of the intermediate compartment (IC) into the array of Golgi cisternae is a crucial step in membrane transport, but it is poorly understood. To gain insight into this step, we investigated the dynamics by which cis-Golgi matrix proteins such as GM130 and GRASP65 associate with, and incorporate, incoming IC elements. We found that GM130 and GRASP65 cycle via membranous tubules between the Golgi complex and a constellation of mobile structures that we call late IC stations. These stations are intermediate between the IC and the cis-Golgi in terms of composition, and they receive cargo from earlier IC elements and deliver it to the Golgi complex. Late IC elements are transient in nature and sensitive to fixatives; they are seen in only a fraction of fixed cells, whereas they are always visible in living cells. Finally, late IC stations undergo homotypic fusion and establish tubular connections between themselves and the Golgi. Overall, these features indicate that late IC stations mediate the transition between IC elements and the cis-Golgi face.

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Acknowledgements

We thank P. Hauri, J. Gruenberg, B. L. Tang, J. Lippincott-Schwartz and F. Barr for antibodies, C. P. Berrie for comments on the manuscript, and Y. Misumi for G95 constructs. Supported in part by grants from Telethon (E.732), the Human Frontier Science Programme (to M.A.D.M.), the European Community (EU grant No. HPRN-CT-2000–00081), the Italian National Research Council (99.00127.PF49 and 99.00133.PF49), and the Italian Association for Cancer Research (AIRC, Milan, Italy). The human GM130 nucleotide sequence data has been submitted to the EMBL/GenBank/DDBJ data libraries with accession number D86427.

Author information

Affiliations

  1. Department of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, 66030 Santa Maria Imbaro (Chieti), Italy

    • Pierfrancesco Marra
    • , Tania Maffucci
    • , Tiziana Daniele
    • , Giuseppe Di Tullio
    • , Alberto Luini
    • , Gala Beznoussenko
    • , Alexander Mironov
    •  & Maria Antonietta De Matteis
  2. Department of Biochemistry, Fukuoka University School of Medicine, Fukuoka 814-0180, Japan

    • Yukio Ikehara
  3. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA

    • Edward K. L. Chan
  4. demattei@cmns.mnegri.it

    • Maria Antonietta De Matteis

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Supplementary information

Videos

  1. 1.

    Movie 1

    Dynamics of GFP-GM130. Time laps images from COS7 cells transfected with GFP-GM130. The numbers in the bottom right corner designate frame numbers and time elapsed in h.mm.ss., respectively.

  2. 2.

    Movie 2

    Dynamics of GFP-GM130. Time laps images from COS7 cells transfected with GFP-GM130. The numbers in the bottom left corner designate frame numbers and time elapsed in h.mm.ss., respectively.

  3. 3.

    Movie 3

    Dynamics of YFP-GM130 and CFP-VSV-G upon release from the 15 °C block. Time laps images images from COS7 cells transfected with YFP-GM130 (red) and CFP-VSV-G (green) showing the fluorescence recovery after photobleaching of the central Golgi area (white circle) upon release from the 15 °C block.

  4. 4.

    Movie 4

    Dynamics of GFP-G95. Time laps images from COS7 cells transfected with GFP-G95. The numbers in the bottom left corner designate frame numbers and time elapsed in h.mm.ss., respectively.

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DOI

https://doi.org/10.1038/ncb1201-1101

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