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Dictyostelium discoideum RabS and Rab2 colocalize with the Golgi and contractile vacuole system and regulate osmoregulation

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Abstract

Small-molecular-weight GTPase Rab2 has been shown to be a resident of pre-Golgi intermediates and is required for protein transport from the ER to the Golgi complex; however, Rab2 has yet to be characterized in Dictyostelium discoideum. DdRabS is a Dictyostelium Rab that is 80% homologous to DdRab1 which is required for protein transport between the ER and Golgi. Expression of GFP-tagged DdRab2 and DdRabS proteins showed localization to Golgi membranes and to the contractile vacuole system (CV) in Dictyostelium. Microscopic imaging indicates that the DdRab2 and DdRabS proteins localize at, and are essential for, the proper structure of Golgi membranes and the CV system. Dominant negative (DN) forms show fractionation of Golgi membranes, supporting their role in the structure and function of it. DdRab2 and DdRabS proteins, and their dominant negative and constitutively active (CA) forms, affect osmoregulation of the cells, possibly by the influx and discharge of fluids, which suggests a role in the function of the CV system. This is the first evidence of GTPases being localized to both Golgi membranes and the CV system in Dictyostelium.

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Acknowledgements

We would like to acknowledge the University of Arkansas at Little Rock for funding this study and providing the resources and equipment to undertake them. We would also like to acknowledge University of Pittsburgh School of Medicine for their assistance in getting this document ready for publication. We would also like to acknowledge Dictybase: it has been a valuable tool, not only for strains and plasmids, but as a source of information and protocols.

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Authors and Affiliations

  1. Division of Nephrology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15201, USA

    Katherine Maringer

  2. Rangos Research Center, Children’s Hospital of Pittsburgh of UPMC, Pittsburgh, PA, 15224, USA

    Katherine Maringer & Sunder Sims-Lucas

  3. Biology Department, University of Arkansas at Little Rock, Little Rock, AR, 72204-1099, USA

    Katherine Maringer, Azure Yarbrough, Entsar Saheb, Sanaa Jawed & John Bush

  4. Biology Department, Baghdad University, Baghdad, Iraq

    Entsar Saheb & Sanaa Jawed

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  1. Katherine Maringer
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  2. Azure Yarbrough
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  3. Sunder Sims-Lucas
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  4. Entsar Saheb
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  5. Sanaa Jawed
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Correspondence to Azure Yarbrough.

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Corresponding editor: Stuart A Newman

[Maringer K, Yarbrough A, Sims-Lucas S, Saheb E, Jawed S and Bush J 2016 Dictyostelium discoideum RabS and Rab2 colocalize with the Golgi and contractile vacuole system and regulate osmoregulation. J. Biosci.] DOI 10.1007/s12038-016-9610-4

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Maringer, K., Yarbrough, A., Sims-Lucas, S. et al. Dictyostelium discoideum RabS and Rab2 colocalize with the Golgi and contractile vacuole system and regulate osmoregulation. J Biosci 41, 205–217 (2016). https://doi.org/10.1007/s12038-016-9610-4

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  • DOI: https://doi.org/10.1007/s12038-016-9610-4

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