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Rapid COG Depletion in Mammalian Cell by Auxin-Inducible Degradation System

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Golgi

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2557))

Abstract

Conserved oligomeric Golgi (COG) complex orchestrates intra-Golgi retrograde trafficking and glycosylation of macromolecules, but the detailed mechanism of COG action is unknown. Previous studies employed prolonged protein knockout and knockdown approaches which may potentially generate off-target and indirect mutant phenotypes. To achieve a fast depletion of COG subunits in human cells, the auxin-inducible degradation system was employed. This method of protein regulation allows a very fast and efficient depletion of COG subunits, which provides the ability to accumulate COG complex dependent (CCD) vesicles and investigate initial cellular defects associated with the acute depletion of COG complex subunits. This protocol is applicable to other vesicle tethering complexes and can be utilized to investigate anterograde and retrograde intracellular membrane trafficking pathways.

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Acknowledgements

This work was supported by the National Institute of Health grant R01GM083144.

Author Contributions

FS wrote the manuscript. IP edited the manuscript. VL supervised and edited the manuscript.

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Authors and Affiliations

  1. Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    Farhana Taher Sumya, Irina D. Pokrovskaya & Vladimir V Lupashin

Authors
  1. Farhana Taher Sumya
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  2. Irina D. Pokrovskaya
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  3. Vladimir V Lupashin
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Corresponding author

Correspondence to Vladimir V Lupashin .

Editor information

Editors and Affiliations

  1. Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA

    Yanzhuang Wang

  2. Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    Vladimir V Lupashin

  3. Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA

    Todd R. Graham

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Sumya, F.T., Pokrovskaya, I.D., Lupashin, V.V. (2023). Rapid COG Depletion in Mammalian Cell by Auxin-Inducible Degradation System. In: Wang, Y., Lupashin, V.V., Graham, T.R. (eds) Golgi. Methods in Molecular Biology, vol 2557. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2639-9_23

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  • DOI: https://doi.org/10.1007/978-1-0716-2639-9_23

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2638-2

  • Online ISBN: 978-1-0716-2639-9

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